So you survived the MidtermCONGRATS! But, now its time to prepare for the final.

Yay! Moving on to defeating the next boss. (^~^)V

Worksheet 4
1. Enzymes increase the rate of a chemical reaction by decreasing the
Activation Energy
a. How does the enzyme do this?
Binding to the substrate, and stabilizing the transition state.
b. What does acetylcholinesterase do?
Breaks Ach into Acetic Acid and Choline
2. Serine Proteases (this will most certainly be on your final)
a. What is the catalytic triad? Give all 3 a.a.s and describe its
function in three steps.
Cleaves scissile (peptide) bonds via acylation-deacylation, and forming tetrahedral
intermediates.
i.
ii.
iii.

Ser - forms a covalent bond w/the substrate

His - accepts H+ from Ser, facilitates bond
Asp stabilizes the (+) His to increase cleavage rate 10,000x
faster
b. What is the oxyanion binding site? Why is it important?
Stabilizes the tetrahedral transition state by forming 2 H-bonds to the (-) charge on
the O of the substrate.
c. What is the substrate specificity pocket? What does it recognize in
Chymotrypsin/Trypsin/Elastase/Subtilisin? What about the nonspecific binding site?
The specificity pocket recognizes specific types of a.a.s
Chymotrypsin- recognizes aromatic a.a.s
Trypsin- recognizes (+) a.a.s
Elastase- recognizes small, nonpolar a.a.s
Subtilisin (prokaryote) recognizes large uncharged residues
The Non-specific binding site assists in positioning the protein backbone via Hbonding btwn carboxyls and amines.
d. Mechanism of Action/Reaction steps: Describe the Acylation then Deacylation steps.
Acylation:

I)

Enzymes OH group goes into active site (catalytic triad) and attacks
carbonyl of the substrate
Tetrahedral transition state forms
Carbonyl reformed and part of substrate is broken off with a new N
terminus; peptide still attached to the enzyme is called the acyl
enzyme intermediate

II)
III)

Deacylation
I)
II)

H2O enters and its O attacks the carbonyl group on the acyl enzyme
intermediate
Carbonyl reforms and substrate leaves with new C terminus
***You should know how to draw these steps as well!!!***

See slides for picture, drawings expected will be like the ones I drew during section
3. What happens when you put lipid bilayers into water?
They form spherical membranes and hide the HB lipid tails from water, allowing only
the HP heads to stick out. If the it was monolayered, they would form micelles which
are also spherical balls.
a. What does amphipathic mean?
When something exhibits both HB and HP properties.
4. Identify the protein.

Chymotrypsin (top View)

Chymotrypsin (side view)

Trypsin

Subtilisin

5. Write the product and label the N/C-terminus after its been cut by
chymotrypsin:
N-CHANYERS-C
N-CHANY-C ; N-ERS-C (notice that the cleavage happens AFTER the aromatic group
and proteins are read from N to C)
6. Compare and contrast Chymotrypsin and Trypsin.
They both contain glycine within their specificity pockets and Oxyanion hole, but
Chymotrypsins specificity pocket contains Ser which prefers bulky aromatic
residues, and Trypsins has Asp which prefers (+) residues.