You are on page 1of 88

Lehninger: Principles of Biochemistry (3rd edition

)
Lehninger: Principles of Biochemistry (3rd edition)

L
R
R
R
R
! R
! R

% # $ $ "
Electrophoresis
% !Sodium Dodecyl Sulfate SDS
% Isoelectric Focusing

# &
'Ms/Ms

" -helix
-sheet
-turns

( Keratin
( Collagen

) $

X-RAY
Lehninger: Principles of Biochemistry (3rd edition)

# " !

Lineweaver - Burk

#

Carboxypeptidase A A

# % $

#&

"( $
"( !Spermaceti Oil )
" &
"

" * $ !Phosphatidylinositols &
" !Eicosanoids &
"+ ) &
"+
"% # Dolichols

& '

" & #

&& & '

& $
ATP

& ! '

+ # $ # $,
+ # # $ ,
% "& & $-) & & # ,
( "& & $ ) & & ,(
Payoff Phase

"
Lehninger: Principles of Biochemistry (3rd edition)

( $ " Pi - "& & $,
( ATP - $ " ADP - $ ",
( $ $ " ,+
( $ ,%
( ATP - ADP - ,

# $ '
Acetyl CoA !I
!II

(+ CoA
(+ $ $

% % !

(% " & ) #

!Flavoproteins
& * & )
#
" & $
ATP
ATP Synthase

% # ! '

)
)$ $
!) & . )
!
&
" & /)

" '

+ & &
% & #
% &

+ ) $ & #
+" Double Helix #
+( DNA
Lehninger: Principles of Biochemistry (3rd edition)

$ ' ) $ $ ,
,&& # & )
, & $ ) $ $ *

, $ & &
' $ & )
, &
)
, )
R $
,

) & )
# ') $ & ) $ # $ ) ,
) $ ,,, &$
,# #) $

, & # , & $ # )

,! 0 ,

, ) $ & ! &

, #$ ) & &
&& #$ #
,,, ) $ $ $ $ )$ ,

) , $ &
& & ) ,! C )
) &$ !R )
& ,
$ # ) ) &
, * # $

, ) " &

) ! & 0 & ) * &
, $
' ) *

( "
CH2—CH2—CH2—CH2—CH—COO-
Lysine: | |
NH3+ +
NH3

(
Lehninger: Principles of Biochemistry (3rd edition)

R
, ) * 0

' ) &
0 ) ) $ ) ) & ,) R
,R

) & & $ # * )
& $ ,
# , # & $ ,
, ) ' $)

LD & &$ $
,& & $ $

D-glyceraldehyde # $ #
,L L-glyceraldehyde D
& & # L-glyceraldehyde # L-alanine
,
,S R # $ #

L '$ ! $
,D L' # # $
D ,L-stereoisomers &
& ) & & ! * & &
,

,L-stereoisomers ) )
,&& # L D # $
, L D # *
& ) #
$ L$ # & # $ ,& !
, # $ $ #

R( !
# , # & &
& ) R # ( $ &
) R , !.pH $ pH $
,! & & ! &

R
, & ) # R
* * 1 !
, &
! ) & , '
, &
, ! $ &
Lehninger: Principles of Biochemistry (3rd edition)

R
) , & ) ) !
, & 0
,! # # ) ! &
& /) $ # ) !
, !
$* ! $ UV 0 !
,!nm - $* & / nm +
, # & & # &

)
# ) # &&
* & . $& $* ,
' # !
= Log(I0 / I) = cl
'
! molar/liter # .c ) # . I0
$& * & . l # . I
! $& . .

R
# ! & R
, ) )
, ' ' $ $ $' #
, ) 0 & $
& $ , & ' ' $
& & ,! # # & & /)
) ) ) $ #
, )

! R
, ) &
$ ' ! ) pH=7 ) )
pKa & $ )
,!pKa=7.59
,) 2 & $ # #

! R
) , ) ' $ ) pH=7 ) )
,

&
, & )
& zwitterions / &)
,!) . !) .
Lehninger: Principles of Biochemistry (3rd edition)

, $ & 2
,pH=pKa & $
, pKa & & ,) && pKa
) & ,pH ) ) ) & )
' !isoelectric point = pI
,! && pKa ,pI = ½(pK1+pK2)
, ! " )

, & .

& N3 &
,N' C' )) & 0 , C3

, ) & )
X! 4 . ' &
0 $
,conjugated proteins ,! .
,,, & ' )
& & * & ,
,) $

0 , & & & )
,

5 # # && ) *
# # * ) / &&
, * & , & * & * &

) && )
& $ & $ $ * ,!crude extract. $
, & ,
) * , # $ ) & * )
& , $ #
, $ 7(NH4)2SO4 6
& / # & & * &#
* , &$ # , &$
, $&
!column chromatography '
! ) # ,, & $ ) ') $
* ,) ) $ & , &
. &$ $ *
, !5 &$ !
) ) / cation exchange chromatography ' $ &
,solid porous matrix $ $
,!high performance liquid chromatography HPLC $ &
$ $ * & #
& # ) # ,1
, ) & / # !

+
Lehninger: Principles of Biochemistry (3rd edition)

# $ $ "
) ) # 'ion exchange chromatography ,
&$ 'size exclusion chromatography ,
, ) 'affinity chromatography ,"

, & * &
,! & . &
! # * &
, $& &$ &$ $
, ) ) *

Electrophoresis !
, ) && &
) , &$ &$ #
,) $ #
*
,) * # & )

3$ * &
,! &$ $ )

, &$ # 3$ &&

V
= /E = Z/f
) .Z & .
0 * & . f .V
!) .E

!Sodium Dodecyl Sulfate SDS
SDS ) &$ &
,!
,& &$ 3$ && && * , ) SDS
, 3$ && ! &
! * , 3$
&& && * , &$* 3$ &&
, & ) &$ * ) $ & &

Isoelectric Focusing
pH & $ 3$ & ,!pI ) # & & *
) # & 3$ # ) * ! pH
pI & $ 3$ ,! . pH=pI / && *
,

,pI & &$ & & 3$ &

%
Lehninger: Principles of Biochemistry (3rd edition)

,) # $ / 55,,) )&$ # ' ) $
0 ) ,& &
& # & ) & $ ,
,)

# &
) * & # &
$ ) ,)
,

$ * ,) N3 # ) )
# & & # $ ,FDNB
, N3 # $ ,FDNB
,& N3 # ) & #

/ !Pehr Edman & ) & *
phenylisothio $ # , & N3
# * , & & & N3 cyanate
,0 &
,! -( '# )
,, &$ /) , & / *

) , & &
&$ ) ) ,
) # & &
, C3 Ang Lys ) ,
& & 1 $
* # # ) & & # ) * # ! &
, &$ & & &

) $
-" / !Mass Spectrometry
,!
0 & ) ) && DNA 0 &
,) & && "
$ ) & , & & & ) * #
, ) $ 0 & ) ) * # # &

# $ ) / ) ) 0 )
, ) 0 ,
& ) .
) ) .
) ) ) .

'Ms/Ms
& ) & 3# $ & & )
! &
' # )
* & * & ') ,I
) & $& , & & $
Lehninger: Principles of Biochemistry (3rd edition)

Mass ) # * &#
,Spectrometry
#$ ' 0& #$ # $ & $
$& & & #* , #
* , & ! . ,& &
, ) !8 #
) 9 # && & ,II
& $ & ,! &
,!&
# * ) &
,! # & & 0
Lehninger: Principles of Biochemistry (3rd edition)

) & & ) & &
) , )$ )
,) & )$ &

'
, 0 / & &
,) & )
* 0 &
,! -sheet -helix
, &

,! & & , 0 '
,
,! -sheet -helix & )$ '
, #
& , , & )$ '
,
)$ #, & ) '
,)

' 0
) / ) 'Super Secondary Structure
,! - - loop ' $& #

& , - ) # / ) 'Domain
,) * & &
domain * domain domain )
,&& domain #
, & domain

,C -C-N-C & " & &
$ ) C-N & & $ X
$# 0 # #, &
,& ) ) )
, & ) ) ) )
O O
| ||
C + C C C
C N C N
| |
H H
) & &
, & # & ) )
$ & C-N
,C -C N-C ) ,

, & $ &
Lehninger: Principles of Biochemistry (3rd edition)

!Psi N-C # !Phi &
+ & # ,C -C #
) * !- + ) ! + ) ) * ) &
& & $
, &
, & $ # )
'!Ramachandran Plot ) & 0$

# # 1 0$ # )
,
, $ $ ) 0$ '

,&
-helix ' , &
' -sheet

-helix
, $ & *
R 1 & * & & # )
,

", & * /! $ (, Å -helix
, =(-60 ) (-50 )< <(-45 ) # !R & ,
, ) #
$ ) & $ ) ) )
, N3 &

# "- #
) ,) $ & NH ) , -helix
0 -helix ) / , N-C
,

N3 C3 / & &
,!*

, , & $ 1 / 4 helix bundle

"
Lehninger: Principles of Biochemistry (3rd edition)

, ,& D & L$ -helix
* & & -helix
,

,! # #) & $ #) 0
, & $ / ) $ &$ & 0

'
! - 0 R ) & ,
, $ $ 0 &$ ,
&$ ) ,"
, "-
'! ) ! ) $ ! ,
) $ $ ' ,i
, -helix # $
, 0 ) ) ,ii
, & ,(

-sheet
# $#$ # $#$ # & & # ,
, & #& # )$
,! ) )

0 ) R
,) $ &$ $#$ #
&
, # #
,) # ) ' *
, # # &$ ' *
,& )

, & )

& , - * &
, ) R

-turns
-sheet -helix 0 * #, & 1 0
* ! * & , /
, +
) 0 ,
, O

& ) ) # -turns
, ) #

' -turn $
,! $ 1 #$ , ,
, ' # & , ,
Lehninger: Principles of Biochemistry (3rd edition)

) ) & ) ! ) '
,! $

, & )$ *
, # # $

$ #) & 0
$ ,Gly Pro, Thr, Ser '
,& & $ $ * #

& )$ * ,) & &
, !

' $ '

$
& # $ &

) &
$& # / Collagen Keratin '
!,,,

Keratin
&
)
) # 0 )
) & ) " &
# & ) )
Cys, Ala, Phe, Ile, Val, Met '
& &

Collagen
$
! ", "
! .X :) & .Hyp Gly-X-Hyp Gly-X-Pro # &
) 3$
)$ $"
) &
Lys &
$ 0 , & $ )$ $
Hyp Pro ; Ala ; Gly "(;

'
, &
# , & # & $
) $ #
,

(
Lehninger: Principles of Biochemistry (3rd edition)

) $ 1 ) $ $
, $

) $
, ) )
) $ , ) & ,Mr = 16,700 / ) ) ) $
, $ &
,Heme - & ) (" && & 0 ) $

,) $ ) $ & # $ Heme
,! # ) ) $ # ) $
* # $ ) 1 $ $ # #
,) #

, -turns -helix $ + ) $ &

, & ) ) , " * -helix
& , -helix # $ ) $ ;
& , & $ * &
* & R ,X )
, $ !

) ) $ , &
,

, -helix & ) $

NH & $ ) $ Pro *
-helix 0 0 Pro ,!) 0 &
, )$

Heme # # ,) $ ) Heme
& His R ) Heme #
, # & # ,O2 %"

#* , $ & $ &
'
* ,) & ,
, & ) &
, - corner - - loops '
* & # -sheet -helix ) & ,
, -helix &
, $ & #& # & ,"
, #) # #
, ) ,
* # #, & ) ,(
/ # , -sheet
, *
Lehninger: Principles of Biochemistry (3rd edition)

, $ * & , & &
'& # / & / # &
, & $ & $
* ,) pH
, & $ *

5& , &
& ) ,S-S $ & SS $ 3# &
$ $, * # ) & $
* , & &
& ) & /) $ /
,
, & &
,!Renaturation "$ #

' ) # $ && )
, # # &
& && DNA & * ) #
,! ) #

1 )
,Chaperonin

$
& # ) & )
*
* * & * ) ) , &
,
, & ' & ) % + &
') & )
, # / ) ,
* '! ) #
, & domains & * * * ,
& & * ,
,!molten globule. * & ' # , ) &
,, &

X-RAY
* # # &
, & ) # & ) ,
' #
,* + , ; ,X & ,
'
) $ *
&
,# (, ; ,!Nuclear Magnetic Resonance NMR ,
'
$) # *
&$ $
,"
Lehninger: Principles of Biochemistry (3rd edition)

& && $
, ) ) $* X

$ Cl– Na+ !NaCl & & $ X
&$ $ !spacing ,
,X & & $

, $ $ $
$ & # & / #
& # ! $& ) &$
, &

' & ) *
$ # $ , # $, &
) , &$ & & & #
/! # $ & # #
,400-800nm $

, - ,( Å $* X * &
# , & X )$ & * ! , - , ( nm
, X
, $& & X $ &
* #& # & / $ &
, )$ $ & & & & ,) )
$ ) & & & #
, $

'X &
* , $ X ) & ) /X $ $
X & ,0 & 0
& * $ ) , &
,!Fourier Transform # # "
, & * & $
#$ ) # & $, # # $ #
,) * & $ & & X & &

) ) $ $ # ) $ )
& $ #$ , $ * )
,) $ & !NMR

,NMR & &$ X $

) & / 5) $ & *
) #, & & ' &$ ) ' $& ,
0 ) &$ & ) , $
,

, $ %%; * ) ) * &
, $ & &$ # pH ' $
& / $ * $ )
, &

$ # $ & $ & * & #
,X $

+
Lehninger: Principles of Biochemistry (3rd edition)

&$ ) & # &
, & $ ,X
' # &$ ) # & ) , & 0
, ) # &$ &
, &
, - #

$ & & * # ) ) '#
,! & & &

# $ * &
,! )
, ) * ) ) #
,& # & ) ) $ # ) #

) $ & E coli /
, (

& * ) / $ * )
& & 0 * ) ! $ $
, & $

,* ) ) .&$

%
Lehninger: Principles of Biochemistry (3rd edition)

'
,! & & ) $ & & )
, & !6-9 M & $ & &
#$ $ * , & #$
,!ER & ) $ $ & # &$ ) $
, & &

$ # , # ) $ & &
, # ) ) $ ! " ;
$ * # # & ,! &
,!
,Heme $ # ) & )

, ) ) $

,!) $ & & ,((Å & ) $
, && ) $) $ $ ) ) $ , && &
,!) $ & # & & ) Heme
# Heme ,30-40Å ) Heme # )
, & His &
) $
# & Heme ) $
,C-N, C-O ' )
!

) $
O

,* # O2 $ ) $
O2 $# 1 *
!

# & & ,) $
,) $
) & $ ) $
PO2 , & & ) ) $

) ) $ ) $ ) ) 0$ )
* ) # ,!88 ( /
, & )

, ) ) , & ) / !tight T
, & ) / !relaxed R
$ & ) ) & ) & /

,# S , &
# & $ & & ) $
, & ) ) $ & BPG $
& ) $ ,) $ T BPG
,

# BPG ) $ )
) T R & # $ , $ & * & , $
,) ) &
Lehninger: Principles of Biochemistry (3rd edition)

$
, # ) $&
/ ) # $ ) ) $ & &
/) $ $ # ) , ) $
,) $

( 5) $
) ( #, &$ & - &
,!T deoxy !R oxy
, !) & ( & #
, 0 # )
) ' # & # )
* # 0 * )& ,)
,! & & 0 #
& # ) ## # $
, $
& & ) $
, &

, , Å &$ # ##

* & ) - & * * ) $
,) $

' & ) $
) ,
& ) # $ # , CO2 ,
+ –
,CO2 + H2O H + HCO3 ' * $
/ CO2 H+ , ) ,pH &
,!His ) ) $ ) $

$ & !) T His H+
& / * & T ' ,) & $ pH
,) # /)
O H O O– H H O
|| | || | | | ||
+
C + H3N — C — C C—N—C—C—
|| | || |
+
O H H O R

N3 , # ) * # N3 CO2
& && pH & CO2 ,R
,T
, # & O2 ) $ #

* '
' $ R T ) / Concerted (MWC) ,
) ) # , ) /R T /
,) $ ,
$, & ) $ / Sequential ,
,
Lehninger: Principles of Biochemistry (3rd edition)

) ) ) & & ' &
,) ) & &

' ' '
, " &
, * $ $
Val Glu / ) & $ 'Glu6Val
* ) ) $ , )
& , ) < /) ) $
,) $ ) $
) $ , & 'Hemoglobin S
' ) ,) $ & # # & S
,
, & ) $ ) $ $ ,
Lehninger: Principles of Biochemistry (3rd edition)

" !
!
) # , $ #
, & pH ,
,RNA ) #

# , ) # # &$
, $
, &$ ) & &$/
$ # !& ' $& & # & #
, &$

' # $&
, & # / subtilisin
C & / !tripsin )
,
, Gly Arg ) & / !thrombin )

!) # Ilu # ' $& , #
$ ! # # Ilu ,Thr
,# * Thr #
, # * # Ilu

E+S ES EP E +P ' #

, ' &&
$ , & $
# # , 0& $
) & # , & $ *
$ * , $
,

, $ ) * / & ! #
& ) # #
& ) / # ,
& ,) ,
,

, & )$ )
, & / # & % 0
# / # # #
$ , # &
, # # , $

* # * & & &
& ) #
,

2 & #
, 2

, # / Km

"
Lehninger: Principles of Biochemistry (3rd edition)

( '
–,G‡ / RT
KBT* e
k = h $

)
#$ ) KB, h, R
!) 3 T
!Ea $ $ =G‡

, $ & 3 &$ $ )

($ + !
K K2
E + S -1 ES - E + P
K-1
' &
[E][S]
[ES] ,ES ,
(K 1 K 2)/K 1

d[ES]=0 '! # ES & ,
dt
, ) , & ) / K1>>>K2 ,"
V = K2[ES] ' K2
, ) ) [E] # # &$ 7S6 # ,
# , K-2 , # # ,(
,V=V0

,& )& &
,V0 * ;

($
K K2
E + S -1 ES - E + P
K-1

V = K2[ES]
K1[E][S] ES
K-1[ES] + K2[ES] ES

K1[E][S] = K-1[ES] + K2[ES] ' ES &
' 7ES6 &&
[E][S]
[ES]
(K 1 K 2 )/K 1

Km = (K-1 + K2) / K1 ' Km &$

[E][S]
[ES]
Km ' Km

[E] = [ET] - [ES] ' ES E #
Lehninger: Principles of Biochemistry (3rd edition)

[ES] = ([ET] – [ES]) [S] / Km '

[S]
[ES] [ET]
[S]Km ' ES &&
[S] [S]
V K2[ES] K2[ET] Vmax
[S]Km [S] Km '

)

V0 = V max [S] / Km Km >> [S] ,
,) & >. #
V0 = V max Km << [S] ,
, & >. # )
V0 = V max / 2 Km = [S] ,"

, # Km

)

, # # $ ) && ( Vmax
'
d[P]
V K2[ES]
d[T]
# $ * 7ES6 $
'!
V max = K2 [ET]
,Km = [S] # V = Vmax / 2

, # && ( Km
& $ ES $ )
Km = (K-1 + K2) / K1 ' ES
,# &
, $ & #

(
Lehninger: Principles of Biochemistry (3rd edition)

,3 pH

, # && (K2
) # & * #
, #
,turnover number Kcat $
,! 2 ) # & &

$ .'

8) # ) ) 0$
, # $ 0$
* $ # # , # # ) 0$
, $ 0$ Vmax Vo
) # * $ #
, &$

Lineweaver - Burk !
)

' &

Lineweaver-Burk Equation

'
, ,
,Y * Vmax ) ,
, Km ) ,"
Lehninger: Principles of Biochemistry (3rd edition)

'
) &$ 1/S * & ) && # ,
, &
,0 $ & & $ # ,

! !
,) $ ) # # #
& * / # & ) * )
, # $
' 0 ) * && &)
& $ # !Transition State Analogues $ ,
, # )
, # HIV # &$ #
$ ! & !catalytic antybodies &$ ,
$ )$ , # 0
# )$ &$ )&$
) * # &$ , #$
, &

#
) $ # ) $
,!inhibitors
& * ,* * # $
* , # !
, #
$ )
' $ , #
/ # / !Competitive inhibitor / ,
, & & ,
# , $& #
, *
) * , # #
$ , # # / km
# & $# &#
, &$ km * / ,
# / !Noncompetitive inhibitor ) / ,
, #
Lehninger: Principles of Biochemistry (3rd edition)

# , # $ & # )$ *
& *
$ # $ ,
$ ) / # Km , #
, & ) ,
,
! * uncompetitive $

# & # * ' ) * #
5 #

!!
# , & $ & # % $ $ &
$ $ , & &$ ## ,! # Lysis ) ##
,!)
) &, ) * & ) & ##
' &
!N-acetylglucoseamine NAG
!N-acetylmurmate NAM

,< &# $
$ ## ,! - NAG NAM ) & # $ * ##
,!PolyNAG NAG ) &# $

, ,) & ,( ##
, , % ## , ) &
, & &
$ ) # #, ( X $ &
& & && ) #,&
# # ,< ? ,! * &
, & ) !heme ) ! )
, # !A,B,C TriNAG / NAG * ) )
$ ,! & ) & TriNAG
/ "& & $ ,
, *
' & /5 # $&
, # A,B,C,D,E,F 'NAG
, A,B,C &
, E,F $
5NAG * ,,* D

+
Lehninger: Principles of Biochemistry (3rd edition)

) A-B-C-D-E-F &# $ $
# '
A-B-C ) &# $ 0 TriNAG ')
, B-C A-B
, &$ NAM 'NAG NAM ) & '
C C NAM NAM & ) C
$ # E-F C-D ) &# $ 'NAM NAG $ NAG &
,!,,,
,D-E ) &# $ ')

# # $ )
' $ ) Vmax $ &

, # &
' # $ !18O & #

,!Asp52, Glu35 ) &# $

%
Lehninger: Principles of Biochemistry (3rd edition)

, ) * & Glu35 Asp52
') ) 2 * &
,) ,) # , # Asp52
,) ) , & # Glu35
,&# $ ) ) ' ) Glu35
,&# $ & * )
) / $ & ) NAG
$ ) , )
, )
!A-B-C-D TetraNAG # E D) & 6-NAG $)
, ##

5D
,! . 2 !)
# ) # , & ) D
,NAG C1 OH ! $ TetraNAG )

)
, ) ) / !
, / $ !
,) &
, & # )
O ( '!sofa
, & , ) $
# ,D ## &
, $ ) ,)

, & )

& ) # ,pH=5 ,pH
( pH ,&# $ && )
( & pH *& , & $
$ pH $ ,
,

)pH !!

'(
Asp52

'( &
Glu35

$ / ,
, )&

"
Lehninger: Principles of Biochemistry (3rd edition)

Carboxypeptidase A A(! ($
, & ) C & ) #
,50Å & #

# # / !) # ' $& &$ & ) & #
, )
# 1 ) , # & 1 )
'
!His69, His196 ) &
!Glu72 $
,
, & ) ) &$ 1 ) &
, # # # #

, $ & ) ## &
) / &
,

, )

' #
, !
,1 ) !OH ) ) & !
,Arg !"

, Tyr248 * $
, # ) $
, ##

!OH 0 & 0 1 $
) Arg127 ) Glu270 )
,

, # # $
0& ) & & # , & &$)
, &
, &$ $ )

"
Lehninger: Principles of Biochemistry (3rd edition)

% $
$
, & & & ' '
,! # & $ # &$
88 &$ #8 * / (CH2O)n
,)

) #( $
,) & & & & & ' & ,
,! ketose ! aldose

! & & & & ' & $ ,
, &# $
, #, & ! & ' & ,"

, & &$ & $ *

) ' $
, & ) & / )$ & !
, # $ !
$! $ , !"
, & $
, & $& ) ) half-life !
$ # & # ') $) !(
, &
,! / ! $ !' ' ' !
) $ ,
+ $ # $ , &) #
, 3$

) , ) $ #
,

, $ $ &

$ ) & & )
) ) ) & & ,
# , *
,

"
Lehninger: Principles of Biochemistry (3rd edition)

( & # &
, ) & ) !
& & ) #
& / 0 #, !Hemi ,
,
,

) ) / ) )
,! ' # ) /
,) & $
,! ) '
, $ ,!) ' & & &
, & & $

$
,# # # $' $&
, &
/) & ) , & &
,! #
& & ) , ) ) & ) & &
D$ ,L D$ ,! ) )
, #
' ) $
D/) &
L/ &

0
, & $ & #
,& # # &$
, ) & $
, ) & / $
, 2n ' # n) #

# / # &
, # $ , )

""
Lehninger: Principles of Biochemistry (3rd edition)

, $ , 0

' &
!,,, # $ , $ & ,
# ) $ & $ ) ) * # & ,
! &
,"
' & ) $ ,

$
! &

,
' $ /) $ ) ) ) ,& )
,!@ )

Palmitate
, ) # )

Oleate
& " # ! % , ,! ) - &
, $ '!&

) !
, / ) /n
' * *
, # ' 3 , $ o

"
Lehninger: Principles of Biochemistry (3rd edition)

& & $ # $$ o
!
, != &/
& & 0 0 , & & ,% &
,

, $3 &/ #
, $ * #

/! ) & , ) $
,) ! $ $) ) !triacylglycerols

$
)
,
$
, & &
$ , & $ # & $
& & $ , # $
,! & &
, & )
) & # *
, & & $ &

/ $ ) $ )
# & , $ ) #
$ ) , &
0 * * ) &
,! &

!Spermaceti Oil )
, & $ $ * &
,! $ /" ) % ; )
, & $ / )

"(
Lehninger: Principles of Biochemistry (3rd edition)

$ ) , " ) 0$ 3 , " ) &
# ) 0 * / 3 $ ) &
,! -" ) )

'
!( '( ) ) / #)
,

'
! & & )
, $ 0 )$

&
, &
'

$ ' /
$ & , $
) ) , 0
,
, ' $ ) &

' $ /
, ) & $ )#$
, # $ , )#$
&& )#$
, $
, & $
) ) / & )#$
) $ )#$ & ) ,
, & $
, & & & & & & /
'&
! $ & &
!sphingomylin= & $ &
$ & & & , & $ '
& $ & & $ ) & ,!B A O '
$ ! & $
$ , # & ,)

)
) # , & $
# ,) $ ) ) & &
& , & )
,) $

, ) # & $ ! $
& & & , & & &&
, & & ,! #
, $ 0 #/&

"
Lehninger: Principles of Biochemistry (3rd edition)

$ , $ &
,

' ( '$
# ! & ) & & & & $
) & (- ;) & '
, * & ) & & ,+ ;

& &
/ # $ , & & # ' #
$ $ * & #
) # ,! $& )
,! !glycosylation $ &
, $ $ ' &
# &
,! #$

* $ !Phosphatidylinositols &
, # ) # #$ &
Phosphatidylinositol 4,5-bisphosphate #$
) )
$ $ $ ,exocytosis *
, 1 $ $

!Eicosanoids &
, # / paracrine &
, & # *
& , & ) #$
) , &
, , & 1 & &
#$ $ !20:4( 5,8,11,14) arachidonic #$ &
,!
'leukotrienes thromboxanes prostaglandins ' & "

'!Prostaglandins ' $
,arachidonic !PG & $
,!prostate gland / $ #$
,!P , PGF !E PGE / 'PG
,, * / PGE1, PGE2
) * cAMP $ ,cAMP ) # $ PG
, PG
# , & # $ PG
) $ $ &
, & $ ! 0$ ,) $ $

'!Thromboxanes $
!thrombocytes & & ,
, # & # &
# ) ) !NSAIDs & &$ &
PG ) # !COX $ prostaglandin H2 synthase
,arachidonate

"
Lehninger: Principles of Biochemistry (3rd edition)

'!Leukotrienes
Leukotrienes, first found in leukocytes, contain three conjugated double bonds. They are powerful
biological signals. For example, leukotriene D4, derived from leukotriene A4, induces contraction of the
muscle lining the airways to the lung. Overproduction of leukotrienes causes asthmatic attacks, and
leukotriene synthesis is one target of antiasthmatic drugs such as prednisone. The strong contraction
of the smooth muscles of the lung that occurs during anaphylactic shock is part of the potentially fatal
allergic reaction in individuals hypersensitive to bee stings, penicillin, or other agents.

) &
Steroids are oxidized derivatives of sterols; they have the sterol nucleus but lack the alkyl chain
attached to ring D of cholesterol, and they are more polar than cholesterol. Steroid hormones move
through the bloodstream (on protein carriers) from their site of production to target tissues, where they
enter cells, bind to highly specific receptor proteins in the nucleus, and trigger changes in gene
expression and metabolism. Because hormones have very high affinity for their receptors, very low
concentrations of hormones (nanomolar or less) are sufficient to produce responses in target tissues.
The major groups of steroid hormones are the male and female sex hormones and the hormones
produced by the adrenal cortex, cortisol and aldosterone (Fig. 10–19). Prednisone and prednisolone
are steroid drugs with potent antiinflammatory activities, mediated in part by the inhibition of
arachidonate release by phospholipase A2 (Fig. 10–18) and consequent inhibition of the synthesis of
leukotrienes, prostaglandins, and thromboxanes. They have a variety of medical applications,
including the treatment of asthma and rheumatoid arthritis.

D( A
'D
Vitamin D3, also called cholecalciferol, is normally formed in the skin from 7-dehydrocholesterol in a
photochemical reaction driven by the UV component of sunlight (Fig. 10–20). Vitamin D3 is not itself
biologically active, but it is converted by enzymes in the liver and kidney to 1,25-
dihydroxycholecalciferol, a hormone that regulates calcium uptake in the intestine and calcium levels
in kidney and bone. Deficiency of vitamin D CH3 C Isoprene CH CH2 CH2 leads to defective bone formation
and the disease rickets, for which administration of vitamin D produces a dramatic cure. Vitamin D2
(ergocalciferol) is a commercial product formed by UV irradiation of the ergosterol of yeast. Vitamin D2
is structurally similar to D3, with slight modification to the side chain attached to the sterol D ring. Both
have the same biological effects, and D2 is commonly added to milk and butter as a dietary
supplement. Like steroid hormones, the product of vitamin D metabolism, 1,25-
dihydroxycholecalciferol, regulates gene expression—for example, turning on the synthesis of an
intestinal Ca2_-binding protein.

'A
Vitamin A (retinol) in its various forms functions as a hormone and as the visual pigment of
the vertebrate eye (Fig. 10–21). Acting through receptor proteins in the cell nucleus, the
vitamin A derivative retinoic acid regulates gene expression in the development of epithelial
tissue, including skin. Retinoic acid is the active ingredient in the drug tretinoin (Retin-A),
used in the treatment of severe acne and wrinkled skin. The vitamin
A derivative retinal is the pigment that initiates the response of rod and cone cells of the
retina to light, producing a neuronal signal to the brain. This role of retinal is described in
detail in Chapter 12. Vitamin A was first isolated from fish liver oils; liver, eggs, whole milk,
and butter are good dietary sources. In vertebrates, _-carotene, the pigment that gives
carrots, sweet potatoes, and other yellow vegetables their characteristic color, can be
enzymatically converted to vitamin A. Deficiency of vitamin A leads to a variety of symptoms
in humans, including dryness of the skin, eyes, and mucous membranes; retarded
development and growth; and night blindness, an early symptom commonly used in
diagnosing vitamin A deficiency.

"+
Lehninger: Principles of Biochemistry (3rd edition)

( K( E
'E
Vitamin E is the collective name for a group of closely related lipids called tocopherols, all of which
contain a substituted aromatic ring and a long isoprenoid side chain (Fig. 10–22a). Because they are
hydrophobic, tocopherols associate with cell membranes, lipid deposits, and lipoproteins in the blood.
Tocopherols are biological antioxidants. The aromatic ring reacts with and destroys the most reactive
forms of oxygen radicals and other free radicals, protecting unsaturated fatty acids from oxidation and
preventing oxidative damage to membrane lipids, which can cause cell fragility. Tocopherols are
found in eggs and vegetable oils and are especially abundant in wheat germ. Laboratory animals fed
diets depleted of vitamin E develop scaly skin, muscular weakness and wasting, and sterility. Vitamin
E deficiency in humans is very rare; the principal symptom is fragile erythrocytes.

'K
The aromatic ring of vitamin K (Fig. 10–22b) undergoes a cycle of oxidation and reduction during the
formation of active prothrombin, a blood plasma protein essential in blood clot formation. Prothrombin
is a proteolytic enzyme that splits peptide bonds in the blood protein fibrinogen to convert it to fibrin,
the insoluble fibrous protein that holds blood clots together. Henrik Dam and Edward A. Doisy
independently discovered that vitamin K deficiency slows blood clotting, which can be fatal. Vitamin K
deficiency is very uncommon in humans, aside from a small percentage of infants who suffer from
hemorrhagic disease of the newborn, a potentially fatal disorder. In the United States, newborns are
routinely given a 1 mg injection of vitamin K. Vitamin K1 (phylloquinone) is found in green plant leaves;
a related form, vitamin K2 (menaquinone), is formed by bacteria residing in the vertebrate intestine.

'Wafarin
Warfarin (Fig. 10–22c) is a synthetic compound that inhibits the formation of active prothrombin. It is
particularly poisonous to rats, causing death by internal bleeding. Ironically, this potent rodenticide is
also an invaluable anticoagulant drug for treating humans at risk for excessive blood clotting, such as
surgical patients and those with coronary thrombosis.

# Dolichols
During assembly of the complex carbohydrates of bacterial cell walls, and during the addition of
polysaccharide units to certain proteins (glycoproteins) and lipids (glycolipids) in eukaryotes, the
sugar units to be added are chemically activated by attachment to isoprenoid alcohols called
dolichols (Fig. 10–22f). These compounds have strong hydrophobic interactions with membrane
lipids, anchoring the attached sugars to the membrane, where they participate in sugar-transfer
reactions.

"%
Lehninger: Principles of Biochemistry (3rd edition)

'

, &
# , 0 # & # &
, $ & # $ &
' & )
& # & !Micelle
,
) # , & # $ & & # !Bilayer
) $ & , & &
,

$ # &$ & #/
, $ * ) $ & , & #

, # $ &
, & 0 $ / !Liposome #
& ) , # * $ !) $ ' $ & & )
,Gel filtration & #
0$ ) ) #
) , # ) & ) $ ,&
# , ) & & / , && #
& ) & , & )
,

,
Lehninger: Principles of Biochemistry (3rd edition)

& & & 0 $ Hewson ' "
, #
, # $ / Nageli & Cramer & ' +((
&$ $, / Overton ' %
,! . :) $ . ,)

, & &

'
,! & # & #
& ) $ ,
, & 0 ,
, $ && " ;

' * ( -+ Å ) ! '
, " Å ,
$ ) , & '
& ,& & & ,! / E.coli & / & &
(; & & : ;/ ) E.coli &
& $/ & , $ ! '
, & $

cardiolipin / , & $ &
, & & ! $ )
,

) & &
,

* &
, $ # # ) $
&
' & #,
,!Lateral diffusion &
& ) # # #
,Flip flop / !&
, $ #

&$ , & ) '
, $ $ & , $ $ , #

& & $ , $
, & #

$ &
,
Lehninger: Principles of Biochemistry (3rd edition)

, 0 ) / $
' $
& * & '
, ) # pH ,

' $ $ , ' '
!Lipid-linked protein & ,
, & ) * * ,
# / !Octyl glucoside $ & )
& $ # ! ) & & # &
$ & & # ,) & # &
SDS , $ & ) $ &) ,
, )& $ & ,) & $ $ &

) ( '$ &
3 ) * !) Cys ,
,!) Ser $ , !Palmitoyl
,Gly N & !Miristoyid & ,
$ $ !Farnesyl $ Cys ,"
,!Geranyl Geranyl
, )
, &
* &* & 1 ,
,!) -7 6 - 7) 3 6 &

1 ' $ ) * * )
, & &
# & ) , < ?
,loop ) 0
, & * $
& & ,
,

) ) & & Bacteriorodopsin )
, & ) , #) ,
, & & #
* & * ,
, ) $ ) & )
,) & ) )& $ &
* &
& & $ &, &
!=G $ * ,
* & * , & &
,) # # - - ) &
,) & & # *

# !Glycophorin ) $)
0 0 * & )
' & ) ) , $ & $ &
, # )
( ) & ) #
# * $ / 5 & ,! ,
,& * ) #
Lehninger: Principles of Biochemistry (3rd edition)

,< – barrels & <

, ;

, , # & ) ' &

& #
,* $ ) # / ,
, $ # )
, * & , ) &
, ) )
, $ ' $ )$
& ,!+ ? # &
)$ ) ,)$ & ) & , )
# $ &) ) & , * & 0
/) & * & )$
, & $ # /
, $ $
# &$ $ ) ) )
, , $ )& ) , $ #

,,, & / $ ,
,
'
/& '
cotransport $ ( /& '
& , & '

$ , & , & #
&/ # # ,
,

,"

"
Lehninger: Principles of Biochemistry (3rd edition)

& '
'
& & & $ / $ & * &
, $ # )# * & $ ,

$ & / #
, & # $ ,
$ )# # , # *
, $ $ !*
$ $ & ) , # /!Catabolism !
,) & $ # $ ,* $ *
,3 / !Anabolism !

,!ATP, NAD+, NADP+, FAD $ )

# & * , # ) )
, )# $ ;, $

,) ) / Lavoisier

&& # $ * ) ,)
, $ ,)

& $
, $ && / & $ / ,G˚
, # / ,G
# ,) # # /
, $ & $ *
,
& ) ,0 ) /
& )
/ && $ $ , $ ) *
$ & $
$ ,
& & # &$ , * $
,
& & #

A+B C+D
D+E F

$ $ ) & $ # ) /$ $
, $ ,55.5M # pH

ATP( $
, !& # $ / ATP
$
, $ & ,
Lehninger: Principles of Biochemistry (3rd edition)

,& # ) &
,

# & , $ & $ &
# , & *
,G’˚=30kJ & $ * ,

) ' ! 1 $
,! & ,
, # $ ,
,ADP3- 0 ) ATP ,"
, $ ,
[ADP][Pi]
=Gp = =G’˚+RTln( [ATP] ) = -51.8 kJ/mol ,(

ATP & , $ & ! . ' $
, ATP . ATP ,Pi ADP

$0 ATP ) $ &
& $ #$
, ) &

ATP # & $
$ * & * $ ATP # &
, *

$ , $ ) & & * # &
* # $ ATP
, # # & # $ $

! (
, $ ) ) , & # ,
, $ ) # # ,
, $ )
=E = ' & $ # / electromotive force (emf) & ,"
=E & , & , & * # ,emf
# ) , # && $ / &&
, & & , & &
) # ) ,* * ) )
, # ) && ,
'=E =G )

) . # ,
# .)

) / $ & & $ ) ) &
,) * #/

(
Lehninger: Principles of Biochemistry (3rd edition)

# $)
C6H12O6 + 6O2 6CO2 + 6H2O
,=G’ = -2,480 kJ $ *
$ ATP & &
) ,NAD+, NADP+, FAD & FMN /
, &
CH2CH2OH + NAD+ CH3CHO + NADH + H+
,! # & ,NADP NAD #
, # ,! # ) 0 NAD &
, # 0 NADP
# # , # # !NADP NAD
, # ,*
, & # # # & #

NAD
nicotinamide adenine dinucleotide . NAD
,NAD+ ) & ) N ) NAD
88) # ,) )
,!& & , ) * ) NADH #

NADP
# # ,C2 & $ NAD
,

, $ #

!FAD FMN Flavin Nucleotides
, #
, #
,
, # )

, $ # B & isoalloxazine ring

+
# * * & & ) ) ) ,
,FMNH* FADH* ) # & , & # &
,H++e- & #
) # , $ # ) ) & ,
,FMNH2 FADH2 ' )
) ) ) &
, $ & $
, $ # *
Lehninger: Principles of Biochemistry (3rd edition)

! '
)! '
' & # $ # $
,!) )$ $ ) ,
,! # $ ) ,
,! & # # ) ,"

! '
* # $ # 0 # $
,ATP
#
5 ) # , # $ / #
# $ * pH ) ,
,
$ ) & # ,
# Mg2+ #$ ,
, #$ # #

# $ $

$& # $ $

# Phosphoryl transfer
ATP
# $
ADP # $
# Phosphoryl shift
& )
# # * # #& Isomerization

# & & Dehydration

# & ) ) Aldol cleavage
&
! &

!preparatory phase ' # $ $
, !payoff phase
, "& & $ # $ * ATP
,NADH ATP * * "& & $ payoff

) ! '

'
Glucose + 2NAD+ + 2ADP + 2Pi 2pyruvate + 2NADH + 2H+ + 2ATP + 2H2O
Lehninger: Principles of Biochemistry (3rd edition)

Preparatory Phase

# $ # $,
,# ' # Phosphoryl transfer ' $
Mg2+ #$ ) # # ,# $ & ATP #
, ) # ATP

, ) & =G ' $

# # $ ,
,# # # ' # Isomerization ' $
$ C2 ) ,# !# # $ !#
,# #& / & $ )
, #$ * ) $

/ $ 2 $ ' !

+
Lehninger: Principles of Biochemistry (3rd edition)

# # ,"
,# ' # ,Phosphoryl transfer ' $
,# ) ATP #

# , # # $ ) $ #
' $
AMP ADP # & ATP # $ # ' ADP ATP
)$ $ ATP # # ,
,!)

"& & $-) & & # ,
# ' ,!# & # & # ' # ,Aldol cleavage ' $
, & # ,! & & 8 $
,# & # #

) ) # , $ & )
=G * & ) ,* * &
,

%
Lehninger: Principles of Biochemistry (3rd edition)

"& & $ ) & & ,(
,# # # ' # ,Isomerization ' $
) "& & $ # $ * & &
, #

, ) # ) ) & * * $
, #

Payoff Phase
& "& & $ # $
* ATP # ,ATP $ "
,ATP ) / &

$ " Pi - "& & $,
NADH ' ) ,!Pi $ & # ,# $ & & "& & $' #
,! $ )

) ,NADH NAD+ "& & $ )
,H+
SH $ & & / # * )
& # , / # )
# & , ) $

(
Lehninger: Principles of Biochemistry (3rd edition)

$ # NAD+ # $
, &

ATP - $ " ADP - $ ",
8 # # $' ,# $ ' #

(
Lehninger: Principles of Biochemistry (3rd edition)

$ , $ $ & ' $ &
) ) & ,=G(6+7)=-12.5 kJ/mol '
,* * * $ & ATP

$ $ " ,+
8 # # #$ ,# $ ' # ,Phosphoryl shift ' $

$ ,%
, & # ,# ' # , & &' $

$ * $ & )$
, # &

ATP - ADP - ,
' # ,Phosphoryl transfer ' $
,Mn2+ Mg2+ K+ & # ,#

&$ $
) &
'

'
/

(
Lehninger: Principles of Biochemistry (3rd edition)

$ '
) & ) # $
) ,# $
,!respiration * CO2 H2O # $
,CO2 O2 * #* $

)#( 1
# $ * /
,Acetyl Coenzyme A
) $ Acetyl Co A /
) * CO2
,FADH2 NADH
FADH2 NADH # /#
* ,
O2 respiratory chain
$ ,H2O
) # * ATP
,!Oxidative Phosphorylation

# $ * *
,
& ) cyanobacteria
, $ 1
& *
,
) &
, # $

Acetyl CoA )I
)
& $ &
# * ,
!pyruvate dehydrogenase complex #
, # &
' &
$ ' Keto dehydrogenase
) ' ketoacid dehydrogenase
,
,0 & & ) &

axidative CoA *
* ,CoA 0 CO2 / !decarboxylation
! 'H- ' ) & ) ) # NADH ,NAD+ NADH
,!
,Lipoate NAD, FAD, TPP, CoA ' * ! #
,!
,thioesters $ $ !-SH CoA
- # & * ! $ $ *)
, ,! ' CoA 'Acetyl CoA

("
Lehninger: Principles of Biochemistry (3rd edition)

pyruvate dehydrogenase $
) !
pyruvate dehydrogenase (E1) ,
dihydrolipoyl transacetylase (E2) ,
dihydrolipoyl dehydrogenase (E3) ,"
&$ $* # $ # $ &
!core # & Lipoate E2 # ,
& $ ) !-SH Lipoate ,
,FAD E3 : TPP E1 ## ,&

' (

,E1 TPP ) CO2 ,
, ) !CO2 ,
lipoyllysyl & & $ #
, # 0 ) * / E2
,!CoA $ )
,CoA CoA E2 $ ,"
FADH E3 FAD E2 $ ,
,NADH NAD+ & ) FADH2 ,(
# # # # ) , # & -(
, # # # & *

! ) pyruvate dehydrogenase $
) $ * ) $ !TPP )
) $ $ , ) * # $ $ *
,

(
Lehninger: Principles of Biochemistry (3rd edition)

)II

$ ) *
$ $ & ,FADH2 NADH
,0$
oxaloacetate CoA ,citrate synthase # & # ' ,
,Citroyl-CoA #
& $ G ,CoA # & * #
* # ,! & . $
$ # & * $ $ ,axalacetate 0$ * #
# & ) * , $
,
,aconitase # & # ' # ,
# & * # ! # & & & cis-aconitate
0$ ,!* OH H isocitrate #
& $ isocitrate &
,!
isocitrate # & # 'CO2 -Ketoglutarate # ) ,"
, CO2 * -Ketoglutarate # * dehydrogenase
NAD+ & ,NADP+ !NADH + H+ $& NAD+
, $ 0 #

((
Lehninger: Principles of Biochemistry (3rd edition)

-Ketoglutarate & # 'CO2 Succinyl-CoA -Ketoglutarate ) ,
* succinyl-CoA CoA -Ketoglutarate ,dehydrogenase complex
) $ NAD+ & ) ,& ) CO2
,CoA succinyl )

pyruvate dehydrogenase & & #
E3 E2 E1 $ # # $, && ) $
& $ & E1 0
,Lipoate TPP, FAD, NAD # $ ' -Ketoglutarate

,succinate CoA succinyl-CoA 'succinate succinyl-CoA ,(
G ,GDP/ADP GTP/ATP ! $ $
succinyl-CoA synthetase # & # ,!-2.9 kJ/mol *
& , & ) $ & &
&$ ) # ,GDP/ADP !P ) #
, ) #
& / !isozymes #
GTP $ # ,GTP ATP
,nucleoside diphosphate kinase # & ATP
'0$
GTP + ADP A GDP + ATP G’˚=0 kJ/mol
succinate ) & # 'fumarate succinate ) ,
,dehydrogenase
FAD * Succinate
ATP ,( * O2 * 0 ,FADH2
,! # ) ' $ * * $#
, & #
$ ) malonate succinate #
,0$
# & ,fumarase & # 'malat fumarate & ,
& $ ,!H2O & L-malate
,D-malate L-malate ,cis trans &
* ,malate dehydrogenase & # ' Oxaloacetate malate ) ,+
,H+ - NADH ' * NAD+ ' oxaloacetate
& ) * * $ # oxaloacetate
$ ) # * oxaloacetate G,#
,oxaloacetate ) & * #

(
Lehninger: Principles of Biochemistry (3rd edition)

& $ $
# & $ * ) ) $ ! #
# $ $ , -ketoglutomate *
, # & !aconitase ' #

! ) $ *
* , oxaloacetate $ 0 ,CoA
,CO2 * &
, $ * & &
FADH2 NADH ) $ *
ATP ,( O2 NADH ,ATP )
,ATP ,( O2 FADH2

$ ) * & $ $
$ # $ $ & ,0$
$ # , $ # !
$& , & $ * # ,
,) ) # -ketoglutomate oxaloacetate
) !amphibolic pathway $ $
) * & , )
- oxaloacetate $& , $ $ $ )
, & ketoglutomate
, # $ $ * # $ Oxaloacetate
,) $ ) $ heme # Succinyl CoA
,anaplerotic reactions & $

# & # CO2 oxaloacetate
ATP $ * & $ ,pyruvate carboxylase
,&

$ ,CoA # #
, $ && * oxaloacetate &&
# & oxaloacetate phospho-enol-pyruvate (PEP) #
& $ fructose-1,6-bisphosphate & # # ,PEP-carboxylase
, # $

biotin ) # * & pyruvate carboxylase #
) avidin ) , CO2
& $ ) biotin
,!) &

$ '
',,, $ &
! $ ' CoA ,
, " ' $ $ ,

(
Lehninger: Principles of Biochemistry (3rd edition)

CoA
# # ,pyruvate dehydrogenase & #
& ,NADH NAD+ : CoA : ATP AMP $
, $ * $
CoA CoA AMP ATP $#
& ) $ ) ) & NAD+ NADH
,CoA ! $ 0
* & ' 0 $
$* $ ATP # E ) # & ) ,
* $ ATP # E ) # ) ,! .
,
, 2 ATP
, ATP $ & 1 ) Ca2+

$ $
# $
, $
) $ $
, )
,citrate synthase & # ' $
CoA oxaloacetate $ #
$ oxaloacetate # ,
# !+ $ )
# NADH NAD+ #
,! $) + $ NADH
!ATP $
# $* , #
& # $ " ,!succinyl CoA
& isocitrate dehydrogenase
, -ketoglutomate dehydrogenase
& NADH & #

# $ ATP ,Ca2+ &
ADP "
,

$
* # $ #
, $ # $
CoA lactate
& ) & , &
& ) $ ATP NADH
# & ) $ ATP NADH
# $ * phosphofructokinase-1 #
,! $ ' #

(+
Lehninger: Principles of Biochemistry (3rd edition)

% !
$ ( !
, # $ $ ) #
& & $&
,ATP # ) $
$ & # $
,ATP
, # $ ATP #
, & ) #
) FAD2 NADH $ H2O O2 # ) #
, & ) *
#* , ) NADP+ O2 H2O )
) ) & $ ) & , $ )
, #

" & ) #
* & # ,
,
* & $ # & # $ ,
$ & * & * & $ &
, $ )
* & # & $* # ,"
!ATP synthase & # # ATP # $
,ADP ) # # & &

$ $ !
&
& & ) 3$ 3$ $ % +
, ) #
& , &
!Mr<5000
& * &
, $
&
# & & ,!H+
,
,# ATP
$ &
,> B
# ) &
) #
,
*
'
, $ * ! -keto #$
& Pi ADP &)
, & & ATP

(%
Lehninger: Principles of Biochemistry (3rd edition)

$ $
, $ ) #
$ & &
'$ $ # FMN / FAD NADP 2 NAD / !acceptors
+ +
Reduced substrate + NAD oxidized substrate + NADH + H
Reduced substrate + NADP+ oxidized substrate + NADPH +
+
# NAD+ #$ & &
, &
,Nicotinamide nucleotide–linked dehydrogenases & # $& $

NAD+ 7'H-6 & ' ) & #$ & & NAD
, & H+

NADPH + NAD+ NADP+ + NADH ' #
NADH , # $ & & $ NADPH NADH
$ NADPH , $ $ $
,! .
* & & NADPH NADH
,!* /* 0 ) $

!Flavoproteins
, 0 # !FAD FMN ) ) $
!semiquinone ) & ) ) ) ) $
$ # ) ,FMNH2 FADH2
NADP NAD ) ) $ & # ,
) , )
, # )

FMN FAD # ) # & #
,

* $ ) ) ) ) $
, & ) 0 ,
) ! $ & & & )
,!quinone hydroquinone & & )

& * & )
0 & &
, & ) & $ $

' ) # * $
,Fe2+ Fe3+ # ,
,!H+ + e- ) ,
, / !:H- & & ) ,"

* & ) ) #
, # )
* ") NAD 0
'
! & ) Ubiquinone ,
' # )
Cytochrom ,
Iron-Sulfur ,"
Lehninger: Principles of Biochemistry (3rd edition)

Q coenzyme Q Q ! ( ' + Ubiquinone
, * & )
, ) ) &
,* & # & ) # #
* !QH0 & ) * & ) Ubiquinone
) $ & !QH2
,& )
& & * & & Ubiquinone
& ) # & &
,
& # & $ $ $ Ubiquinone $
,

# Heme # & $!
, # Fe2+ ) Fe3+ ') ,
$ , & / a, b, c " &
, " #
b 560nm 600nm a &$ $ *
,c 550nm
Heme , * # b a Heme
,Cys * & c
Heme # & #
, )
, & $ c b a$
) c ) &
,
succinate NADH & * # *
$ # Ubiquinone * &
,O2 $ 0

& # &
'
NADHC Q C cytochrome b C cytochrome c1 C cytochrome c C
cytochrome a C cytochrome a3 C O2,

# # & # &
, # #
) ) / & ) & O2 # & ) #
) ) & 0 0 # $) ,O2 )
, * & ) * &

#
$ * $
) $ & & )& , # &
,! # &

NADH / & Ubiquinone # II I
Ubiquinone III ,!II succinate !I
,O2 c & 0 IV :c
Lehninger: Principles of Biochemistry (3rd edition)

!NADH Ubiquinone oxidoreductase $ ,Ubiquinone NADH I$
(FMN ) Flavoprotein & &$ #
, $ #
, L
')& #
, & ) :D NADH !:H- & & ) $ ,
$ E , # $ & ,
, # &$ & NADH )
$ & *
) * & ) ' #
,! * )) ! &&

' ) * # * $&
NADH + 5H N + Q C NAD + QH2 + 4H+ P
+ +

. & .N : ) . & .P

,EEE E !DH #
, ) H+ $ ,D # !)

,amytal, rotenone, pericin A ' E

!Succinate Dehydrogenase $ ,Q Succinate II $
I ) # , ! %
D C , ! $ # # FAD $
Heme , "
,! ) ) Ubiquinone
FAD 2Fe-2S # , B A
,) Fe-S # ) FAD # .

* & #$ & &
,II * & * )

Dehydrogenage Acyl COA ) # Acyl COA <) )
) ) ,# $ & & FAD
,ETF: ubiquinone oxidoreductase ) !ETF
" $ ,Ubiquinone # & * # #
# & triacylglycerol $ )
# # , $ $"# #$ $ & ) # $ dihydroxyacetone phosphate
& )
# & * / # $ & & Acyl-CoA # $ & &
,Ubiquinone
Lehninger: Principles of Biochemistry (3rd edition)

# NADH # $ #$ & & " $ &
* # & , & *
!QH2 Ubiquinones 0 , # Ubiquinones
,III Ubiquinones

,G-3-P Acyl-CoA ' )

!bc1 $ ,cytochrome c DF III $

' , # & 'bc1
cytochrome b (2 hemes)
Fe-S protein (2 Fe-S centers)
cytochrome c1
/ !E &
'* * , )
QH2 + 2 cyt c1(oxidized) + 2H+N C Q + 2 cyt c1(reduced) + 4H+P
/& ) Ubiquinone Q $
* & $ # &$ / c,c1,b
QH2 ' & $ 0 * * ,c
) ) c , # c Q )
& IV III ) && Heme
, # # )

!F G # G cytochrome c IV $
, )
) & " & 8 & &$ & # #
, & * ,Mr=204,000 ,! )
) & &
) & a3 a Heme ,CuA & $ &#
Heme a $ &# & CuB Heme ,CuB
,O2 Heme a3
' &
Cytochrome c C CuA C Heme a C Heme a3 C CuB C O2

! * #
) ) # $ $ , H2O O2
# $ & 0 * ) # !
,III I * & &

' # & # *
4 Cyt c (reduced) + 8H+N + O2 C 4 cyt c (oxidized) + 4H+P + 2H2O

& & ) # # O2 #
# $ ) & 0 & #
, $ # & ,

Antimycine A CO !CN- & )

& $
, & $ ) $
'* ) * & NADH
NADH + H+ + ½ O2 C NAD+ + H2O

"
Lehninger: Principles of Biochemistry (3rd edition)

O/OH $ # NAD/NADH & $# ,! $ . $ & #
' E’˚ , G’˚=-220 kJ/mol NADH/NAD / $& , $
,! & $ , E’˚=1.14 V
,20 kJ/mol $ )
, / 200 kJ/mol

) # $
' G ) $# ,
'3
'333
'43
H2O ! $# 0
,VI

' * )
NADH + 11H+N + ½O2 NAD+ + 10H+P + H2O

$ & # # & $
The # & $ $ , $ #
' Proton motive force
& & # # & $ ,
, ) '
& * ) & $ ,
, &$ 0 &

ATP ! $
5ATP $ & $# &
&$ $ Proton Motive Force $
& ) # ,50kJ & ATP & ! $ # 200kJ
,* & )

' &
* & # & $ # &
# ATP # &
, *
, #

' ) *
ADP + Pi + nH+P C ATP + H2O + nH+N
Lehninger: Principles of Biochemistry (3rd edition)

! ) Pi ADP && &
' " ) )
,! * # ) ,
, O2 ,
,# ATP ,"

,Pi ADP $ ) ATP # )
#) & ATP # ) $
,ATP # O2

'8 ) * $ &
# & & ATP #
ATP ,ADP * ) O2 && &
,* O2
)$ & $ $ ) &
, & ATP synthase digomycin
* & $ ATP # $ # )
,O2

* ATP synthase $ & ) $ $
, * 0 & ATP #

# , & ATP # #
!) $ & ATP synthase * & *
* &$ * #
, &$ & $
$ &
,O2 NADH & $ # & $
#* * $ , # #
, # / O2 &

& ,) ) #& $
) # ) & $ &
, * & / ATP # * O2 NADH
) , & $ & )
) & ) ,FCCP DNP
) !88) & ) , & * &
, & $ # * )

8 / !F1 ATP $
, # ATP & )
,ADP Kd=10-5M Kd<10-12M &
,ATP $ &
,Proton-motive force $

)ATP( 1 !
,,,3 CO ) & ' ,
DCCD ) $ ' # ATPase ,
FCCP DNP 'ATP # ) ) & ,"
,ATP ADP ) & , & ADP ,

(
Lehninger: Principles of Biochemistry (3rd edition)

)ATP( 1 ! $
'
ADP ,) # # ATP && ) ,) # ) # && & 0
! ) , ) Pi
, ) ' & )
,) & ) $ !CN & )
# ) & , ATP # 'ATP #
,ATP
, ATP $*

'!
ADP + Pi ,ADP + Pi
!& DMP ,) !) $ ,
, $ ATP # ) /& &
, $ & &
# ) * * & & & &
ATP # ) ) & $ ,
, * & $ & & ,! *
, $ # # # & * #

)
& # ,NO2 OH )# 'DNP (Dinitrol phenol)
& ) & # ,!OH C O-
,* $ ,
) $ & ! # 'FCCP
, &

5 & $ & * & ATPase
* ) # , ) & pH=9 & *
,ATP # &
, # !ATP # ) $

ATP Synthase
# # & ,F & ATPase
# &$ # , ATP synthase
Pi ADP ATP # &
/! ! & #
V $ # ,
& ) / F1 '
$ ) Fo o , / Fo !
,Oligomycin
& $ * O2 NADH # F1
& Fo ,
,ATP # $ & Fo ,

, # ATP ATP # & $
) # & $ * ) ATP
, # ATP ) & $

, ?3 3 # $ ( & % & F1
) # 0 ,ATP # < &
) # <
Lehninger: Principles of Biochemistry (3rd edition)

) < ,I H & ,<
,ATP/ADP
#, & ) F1
,

& & c & ,a,b,c & " Fo
< , &
,& ,?1
,F o F1 ?1 < &
, $ ) #c & !c & Fo &

, ATP & < & *
, Pi ADP ADP < < &
$ # # &
,ATP # # ,ATP < / &
& 0 ,ADP+Pi ATP ' #
# ATP $ & / ATP & <
& # 0 , # #
, # ADP <
# * & &
c & $ $ Fo * &) #, # Fo
* * &
,
& # ?"<" & # * &
, H b2 &
1 < & $ J
, <
< # < &
ADP < <
/ ,ATP < &
ATP " # * &< &
, #

'! &
O = open L = low effinity T = tight

' ) ATP # FoF1 & & ) $
* ,< ? & ) & His &
) Fo ,F 1 * & Fo
, J
# / &) $ $ ) &
ATP , ) & ) / #
, & $

'!F1 0
) * ,) * & ,) ) ATP
,!ATP & ) && ) & ) * # &
, ! -" ) '
5ATP # & $ * , GK0 # # & &
# ,!ADP $* # & ) # # ATP , )
& ADP , # ATP $ , ) G
, $ ) * ATP
Lehninger: Principles of Biochemistry (3rd edition)

,ATP ) & * # )
,ATP # & ) &
, $ ATP /

ADP/ATP ' , ADP # * & ATP )
,!ATP # ) $ #) ,translocator
,! & ) $

,Succinate ! ) ATP
8 $ oligomycin ATP
,) * &

"/6 * "- &

)! ' ATP( 1$

+
Lehninger: Principles of Biochemistry (3rd edition)

# ! '
* $ * )
, # $
& , # $
$ , # ! & # $
&
,)

$ ) # & #
, # $ #
& ) $ ) $
, $ &
0 /& # & $& / )# )
, )
) && $ &
,) & # ) )$ $
, # $

' & ) & # $ *
) &1

&

&
,,,* $&

, && $ & &

&
& # ) # $
, ) & & ,
, $ & # #
,!88 $

*
$ & & # & # )
, $ $ ,

*
'
$ ! ) ,
) $ * ) ) $ * * & & ) ,
, $
,) & * $ /
* #
) # ' , $ $ &/ $
, $ * / # &
) ) ,! # &. $
,! . *

%
Lehninger: Principles of Biochemistry (3rd edition)

/ & $
, $ ) * / ,! &

' $
#, $ 2 $ ) ) ,
,!) ' $& ,,,3
, # ) * # ) ,
# & ) ) G ,"
, #
, # ) ,
& $ * & ) ,(
,)
)$ !mRNA DNA #$ !) $ & ,
, $ & , &

* !
) $& ) , $ $ # * $
) $ # $ ,)
, # $ ) & #
,) & # $ # & 0$ # ) # )
') $&
,TRH / ! / #
, # !88 )
, & / 0$ * & # /5 * &
,! ) & # ) & $ &
'radioimmunoassay (RIA) )
)&$ & ) #) ,) )&$ ) #
& ) ) $ ) ,
, !) &
,) ) ) )
) & ) ,(picogrem??) pg $
, )

) '$ #(
& ,
, "-
, # ' $&
& $ ) # #
* & , 0 &$ 0 # , ! #
, 0 *
, # 1

,
, ! & . )# #$
, & #
, 1 $
,
,L-DOPA ) & !) ) & ' $&

& ,"
, &
,carriers & # & #
Lehninger: Principles of Biochemistry (3rd edition)

, $ # #
, & & $
,
,) ) $ ' $&

)
, & $# $ <
, $ $ '

)
) & '
$ # # $ # $
)$ $ # )$ $ #
)$ $ )$ $
# $ # $
CoA
, , #

)$ $
, & # $ ?
, &

)
) & '
, , )$ $ )$ $
$ )$ $ #
# # $ # $
& $ , , & #
& # $ # $
# $ #$ $

!) & . )
, $ 0$ & stress & &
, $ )$ $ &
,) )$ $

' $ #
$
O2 &1

)
) & '
, , )$ $ )$ $
$ )$ $ # )$ $ #
#$ $ # $
Lehninger: Principles of Biochemistry (3rd edition)

) !
) .'
, $ & $ '
# ' &
& )# 'portal vein
, & $ & )# '&
,!# $ # ) '

&
, & $ & )# &

' &
# $

)

$ ! (
# 0$ $ * ) $ / 0$ &
& ) ) #, # &
, $ # & /
,# $ GluT2 !& . # $ &
) * ! # $ # # $ # $ ) #
,# $ & Km ! ) # $ !
Lehninger: Principles of Biochemistry (3rd edition)

'! # $
* & , & # $ # # $ & ,
,4-4.5mM . & # $ #
,)$ $ # ,
& L $ L # $ ,"
, $
$ 0& & /8
,# $
,) # !" 0 A ,
# (# # ,(
,NADPH &

# $ )) & * # $#
# $ $# $# , )
,&

. 1 : . 1 /8
,! $*

! +
, & & # ,
, & * & ,
, )# ,& & # ,"
, # $ CoA & ,
, $ * &
1 * ) ,(
* , & & ) ! 0 !*
, #$ $ # $

! +
' $ & ,& & ,
$ $ $ CoA !<-oxidation ) ,
, $ & ) ,!&
$ $, & $ * CoA 0& ,"
, $
,CoA # ,
, !) . & & $ & ,(
,! & / !) $ & ,

'
$ CoA # * & # CoA &
, *
,D-<-hydroxybutirate ) ' $&
, ) $ &

& /)
& # & $ & , & $ $ ! & )
, & &

, & $ # (; ,) 0$ (; / 0$

"
Lehninger: Principles of Biochemistry (3rd edition)

) ) ) /) & $ ) ') *
cAMP # ) & # G)
,!perlipin ) $ 0 ) & #
,) & 1 & ) ) )
, $ * * & )

# # & & & $ & &
, & )

) )
& $
,1 )$ $ & # $

,) # ) & $# ATP
, $ ) ) & $ '
pH & & )$ $ ' $
,) $ #, $
ATP & ) $ & & 0 & )$ $ &
,10-30mM ' $# # # , # * & ADP

' $
,) )$ $ # & * & ' ,
& & , $1 ' & ,
, Heme
, & * ) / & ( ;

' $ $ ,
$ , &
, $ ,
&$ ,!pg & ) '
,mg & ,! g
, & # ) &

'& $ ,& #
, )$ $) ,& $ & # $
, $ - ;) # ,& * & $

, $ & $ '

, & &
Lehninger: Principles of Biochemistry (3rd edition)

! ' ! ! + '
,! )# $ ! & &
mg
, $ $ / 0$ $ $ , -% /mL ) 0$ )#
,

,) # # #
# ,0$ # & &
,) < )$ $ ? ,H < ?' $ / $

!Glut2 # $ <
) & $ # $
# # $ , #
, $
# ATP # $
,)$

)$ $#
'1 * # *
)$
) # # &
# &
,
& $
& / #

, $
$ & )

)&
)& ) # #
)& # !* & #
* #/ $ *
,)& # & , $

& ) , ) # $ $ 0$
,&

$ $ $ )$ $ # $ &
) $) ) ,!) & ) & * &
, &$ # $

! )$ $ # & # $ &$ ' $ $
, # ! $ &$ $ / # $

) #* , & 0$ & # $
) # $ )$ $ & & # $ & & ,) $ $
, $ & )
, $ ) / #$ $

(
Lehninger: Principles of Biochemistry (3rd edition)

,0$ & *
,# $ )# / $ $

!& # 51
! $ & $ *
& * #, #* , #
!0$ $
#$ $ *
,
$ , * & $
) * &
, $ Co-A ,Co-A

$ & # $ ' $
# $ * # & #
& ) # , &
) $ $ )
,

* )
#, # $
* & # $ * )
* 1 $ * # $ # ,
# * $ , #
, &

/ $ , # &
& $ & / )#
& , # & ) ,)
$ , & & & # & /
# # # # & ,0$ # # # & /# $
$ & #* , # &$ / 0 &
# & ) ,)
, # & & ## $ ,) 0 & & )$

!
, $ #
# Km * ) , #
# & & # ) , $ #
& ) $, & & # * / & & #
,) # *

, )
$ * #
, &
, & # $
& & &
,! / $ &

& ) $ & , # $
, &$ $ #
, # $!
,! & # ,!Ter, Ser, Thr, His
Lehninger: Principles of Biochemistry (3rd edition)

/ Tyr ) $)& / & Tyr ) & / &
, #

& # )$ $ # )$ $ #
$ & * # , # ) #
, & # ,

, & ) / # # &
,! )$ ) # / !
, # #
) $ $ & / # # ) / $
, # # *

,! ,
$ & , $/ # &
, /
Lehninger: Principles of Biochemistry (3rd edition)

'

) $* ) # &
, / & ,)

& &
) ) /$ ,
!ATP, GTP $ ,
!CoA, FAD, NAD # ,"
secondary messengers !cGMP, cAMP $ ,
) # $ ,(
!rRNA & ,
& # & ,
* & & & &
& ) )# & # ,
$ $ & ) )# ) , $ & )
,!A $ & # ) $

, $
, .)& : .)

,!G ) $ ! A ) & ') $ "
,!C ) # !T ) !U 'RNA ') $ #

, & ) $ #
& !DNA ) $ 1 &
& & ) ) & & ,!RNA
,!DNA

+
Lehninger: Principles of Biochemistry (3rd edition)

& ,!C ) # !U ) & !G ) # $ !AMP ) # & ' &
& & DNA $ * RNA
, 5- Methylcytidine

& #
$ * ' " ,
,
, / ,
# conjugated molecules , ,"
,! ) & # #
,
, & # : & ,
,exo, endo ' $ # ,(
, $ &# $ & ,
&$ # & ,! $ &) ,
& $) ,DNA & / !Stacking Forces
, &$ # $ /& # 0
) ) * &# $ $
& * , & &
, $ ) & &$
, ,+
* ,UV DNA 260nm $* # 'UV ,%
, 0$

&
' &

Salvage Pathway /
, & * # # & &
& $& & #
, ) & #* ,!& #

De Novo Synthesis /
' # & ) & & #
,NH3 CO2 (#
#* )& ) ) & ) $
) ,# ) ) / ,!

%
Lehninger: Principles of Biochemistry (3rd edition)

# orotate )& ,* & * # $ &
,) $ # & )& #
') & ) & * &
, # '(Phosphoribosyl pyrophosphate) PRPP ,
, & ) $' )
, & * ( ' ,
Adenosine-5’-monophosphate (AMP) ' ) &
) & ') $ & ,Guanosin-5’-monophosphate (GMP)
,) $
' & *
/ PRPP C1 ) $ & * ) !
) & , & /) # ( )
, #&
& ATP ) ,) $ " !
,) $
,C !"
,ATP & / ) ) $ !
,) $ ATP ,) / #& $ & & !(
,) * " ) &
, ATP & / & !
, #& & !
, 0 !+
) & !%
, ATP , &
,* ATP ( *
N10- /) & ) !
,!) " $ formyltetrahydrofolate
,IMP & $ !

, # $ IMP #

)& ,) * )&
# & # )& , PRPP
,NH+4 CO2 # ) #
, & &
/ A, G, C &# , & # # RNR #
# # , # # ,U &
# &, & & & & &
) ) # ) CTP UTP )# ) , & balance
, ) $ & )# DNA # , &

$ & * # ,!C U )# &
,) #
&$ :RNA ' # & +
,DNA
& & / / & & $
, & + ) ,rRNA tRNA

# * PRPP )
# & !Phospho- -ribosylamine ) # )
, #

+
Lehninger: Principles of Biochemistry (3rd edition)

) $ / & & ) * DNA #
&# & , # ) # )& * , #
,
#* , &# * & # &
,ATP + AMP A 2ADP '* # & & # ADP AMP ,
$ ATP , # ) * & $ # & ATP ) # * ADP
# , # &# # & & &#
# !# & # * * &
,) # & & & ) # ATP ATP + NMP A ADP + NDP
, ) * ATP ADP #
# & &# # & & &#
* ) & # # ,NTPD + NDPA A NDPD + NTPA #
# ,!# & # !) & )
, !Acceptor !Donor
, &# $ # ATP &

& )# & / * , & C2 & # & &
# & # ,!dADP & )# & & 3 # !ADP
& &# # # , # ,ribonucleotide reductase
,thioredoxin ) ) * & DPH $ ) $# &
, & &# / SH $# # ) #& )
# & & & &# # # ) &
,# &
) & $ ,) $ & # # & & # $) & $
, & ) &

$& # * ) # & &
, &
$ R1 & ,R2 R1 & & #
,R2 R1 & ) # , $
, & # R2 ! & SH R1
# , # & # Fe+3 $ R2 &
$ & & * $ & &
, #
, "3 & & ) ,
, & ) & & & ,
, ) ,"
, & ) & ,
," & ) # ' * ,(
, & $ # ) & ,

' $ $ R1 &
dATP # ATP # * $
,
ATP, dATP, dTTP, dGTP ' $ $
,CDP UDP # dATP ATP
, ADP GDP # dGTP dTTP
0& ,) & & # $ dATP
# & $ dTTP $ & ) )& &
dATP $ ADP # dGTP $ ,GDP
, #

+
Lehninger: Principles of Biochemistry (3rd edition)

dUTP dTTP
,!U !T ) DNA
# UDP :dCTP # CDP ,& # & ) & *
,# & & # * dTMP dUTP :dUTP
,DNA T * U # & ) &

'

) $ & #
# $ , # # #
, # # RNA # DNA , & /
& , * / ) # # #
) ) # "
, & & ( & & ?
# ) & #
, !PPi
( & ) ) " & ) & #
) ," & 0 ( & & ) & &
) 2’ 5’ ) ) ) ,5’ 3’ ') &
,#
,!25kJ/mol * # & #

# & template * # , 'Template
'0 &
,!GAC ) # ) $ !AAT ) ) &

#
, ,5’ 3’ & ,
, & & ) ,
,!PPi $ & ,"
, & ,
) & 0 # 0 ) !template & ,(
,

,!# & & ) & & DNA
# & , ) PH $ RNA
, $ & ) & & ) DNA ,
, &$ # ) $ DNA

pH=7 &
& ) , & # & ) , &
&& &$ , # # &$ $ #
& ) & $
, * , ) & $ $ #, &
, * & ) $

Stacking Forces
/ , & #
& $ $ , &
/ # # , &
, & & )&

+
Lehninger: Principles of Biochemistry (3rd edition)

, " G C # # T A
& * $ & , & )
& ' & , ) $ &
,) & )
$# 0 )& ) # # & ) &
/& / ,!10.8Å.
, &
$ ) ) * $ &$ & $
5’ 3’ ') & ) ,& #
3’ 5’

) ) * * ) )
,
) RNA DNA $ &$ & ) ) &$ &
, &

& # Stacking Forces &$ &
double helix #,
, &
minor ) # & &
double , ) & / !major groove &$ !groove
$#) ," J $# ,20Å & helix
,3.4Å

) B-DNA &
, J +J $#) # ,
!propelled # # ,
,
$#$ # # , GC / ,"
,!88 Z-DNA

Double Helix #
, $ & $ pH 3 , $ & RNA DNA
&$ UV &
/ & 1 & * ! & # / &$ & ,
, ,
, 3 / tm
& &$ , &$ ) & $3 * G C
,
template & ,! PPi in vitro $
&$ , $ & &
,PPi #
, & # & ) * & # #
* DNA DNA # , DNA
& #) , ) UV ,
/) # * 0$ & ) ) # # ,& UV
, &$

, DNA / B-DNA
&$ * & ,B & , DNA / A-DNA
&$ ) ) , ) )
&$ ) ,double helix ) ,
,!10.8Å 11.1Å &

+"
Lehninger: Principles of Biochemistry (3rd edition)

,! 0 RNA T A G C $ #
, " J #
$ # ," J & $#
0.34nm $#) : 2.0nm. ' &
DNA , * ) DNA
&$ * 0 & DNA ,! $ &$
, 1 & * & &$ $
'
* & coil & , * & 'Super Coil o
super coil # ,
$ # / DNA $ ,!topoisomerase '
, )
&$ ,!Positive super coil 'Topoisomerase II o
& * # ,* ) ) # # ,DNA
&$ & DNA # , &$ & $
ATP & &/ , $ # &
,! #
') negative super coil positive super coil ) & o
,
) ,

)
* & &$ DNA * # 'II ! !
, # , & /&
/ $ &$ # * DNA
,
&$ !nick & &$ 1 # 'I ! !
, #, & $
, ) & #
nick , &$ * & ) ) * & 0 'nick
& OH # , "3 & 0 & &
, 0 & & & )
, $ relaxed DNA

Linking Number
&$ & & $ , & &$
# # , $$ ) $
,
, ,( ) , ,
) DNA * * # $ !linker )
, DNA # ,! #
) ,* $) 0
,scaffold ) $ & &

double helix topoisomerase histones fibers of histones
loops of fibers scaffolding

, & * #, & # * MNO

DNA & # 5DNA DNA # ) $
5 & # #

+
Lehninger: Principles of Biochemistry (3rd edition)

# & , &$ )# * $
,1n DNA & ,
) # , 1n
,! ) & DNA #
* DNA ,)$ $ * $
, / DNA
,, )$

,!mec 1 )$ )$ / #
,)$ $ * ,DNA #

,* # ) $ ')
, $ mec 1 )$

DNA
, 0 # ,origin of replication '& &
' #0 , &$ ( 0 e coli
& tandem , & " # $# " ,
,!0
,) & # , # $#% ,
,) & ) ,

# $ ,% 0 ADNA
, # # & , &$

+(
Lehninger: Principles of Biochemistry (3rd edition)

+
Lehninger: Principles of Biochemistry (3rd edition)

+
Lehninger: Principles of Biochemistry (3rd edition)

++