ENZYMES IN GLYCOLYSIS

Description Activated-Inhibited
HEXOKINASE Present in Km= Vmax Maintan Hemolytic Phosphorylate Glucose-6-phosphate
all cella 0.1mM lower intracellular anemia monosaccharide
[glucose]
GLUCOKINASE In liver, B Km= Vmax Maintain blood Diabetes Phosphorylate Insulin –during well fed state Fructose-6-phosphate
islet cells 10mM higher [g] melitus glucose
of
pancreas
PFK1 Under complex allosteric regulation Fructose-2,6-Bisphosphate, ATP, low pH, citrate
Function as tetramer w/ 2 states (Relax + Tense) Amp, Adp Shift equilibrium to T (lower
Shift equilibrium to R (higher affinity F-6-P)
affinity to f-6-p
PFK2 & FBPase2 Control level of F-2,6-B PFK2 – F-6-P
Balance is determined by the activity of cAMP-dependent kinase (CDK)**
PYRUVATE KYNASE Function as a tetramer (tissue-specific isoenzyme) F-1,6-B ATP (muscle)
AMP
Alanine (liver)
Acetyl coa
Phosphorylation via cdk
 Inhibited by Glucose-6-phosphate

Phosphohexose isomerase

** WELL-FED STATE Fructose-2,6-bisphosphate

High level of insuline > AMP
decreased cAMP > decreased ADP
level of active Protein Kinase A > Aldolase
PFK-2 dephosphorylation > PFK2 Triose Phosphate
activates, FBP2 deactivates
Glyceraldehyde-3-Phosphate ATP
Formation of F- dehydrogenase Low pH
Citrate
2,6-Bisphosphate Phosphoglycerate Kinase
WHAT HAPPENED?
- F-2,6-B increased, activates
Phosphoglycarate mutase
PFK1
- glycolysis increased, inhibit
glucogenesis Enolase
Fructose-1,6-bisphosphate
AMP

** DURING FASTING ATP

High level of glucagon > increased cAMP > activates PKA > Alanine
PFK2 phosphorylation > PFK2 deactivates, FBP2 activates Acetyl CoA
FORMATION OF F-6-P Phosphorylation via cAMP-
dependent kinase
WHAT HAPPENED?
- F-2,6-B decreased
- Glycolysis decreased, promotes glucogenesis


COMPLEX I
• Flavin mononucleotide, Iron,
Ubiquinone (CoQ)
• H+ and e- trf from NADH to C.I
• H+&e- trf to mobile electron
carrier CoQ--> CoQH2
• CoQH2 carry e- from CI to C.II to
CIII
• loss of H from NADH regenerate
NAD+ (TCA Cycle/Glycolysis)
• Pumps 4H+ out
• 4H+/NADH
• Rotenone & Amytal
GENERATION OF ATP (OXIDATIVE PHOSPHORYLATION)
-Proton dependent ATP synthase
-Use proton gradient to make ATP
-Proton pumped thru channel on enzyme
-Intermembrane space --> Matrix
-4H+/ATP
-chemiosmotic theory
- E- flow thru a chain of membrane bound carriers
- Free Energy made by downhill e- flow coupled to the
uphill transport of proteins across a proton-impermeable
membrane
- Create transmebraneelectrochemical potential
- Provides E for synthesis ATP
-Inh: Oligomycin, and DCCD stops proton uptake
COMPLEX II COMPLEX III COMPLEX IV
• CoQ, riboflavin, FADH2 binding
• Q cycle (Cyt.b, Cyt.c, Rieske
• Cytochchrome oxidase
site Protein)
• FADH2 trg H+ to e- to CoQ-->
• 4e- from four cytochrome c
CoQH2
• Cyt c (contain Fe3+/Fe2+) passed to other e- carriers
• e- trf from CoQH2 ->
• • 4e-+4H++O2->H2O
cytochchromes b/w CIII and CIV
• Generates E from e- trf to pump
• Regenerate FAD 4H+ from matrix, increasing the • Pumps 2H+ out
H+ gradient
• 8H+/NADH • 10H+/NADH
• 4H+/FADH2 • 6H+/FADH2
• Antimycin A CO, CN
 STAGE 1 : ELECTRON TRANSPORT § No ATP synthesis
Electron transport drives pump that pumps protons across
membrane
Intermembrane space high [H+]
Matrix low [H+]
STAGE 2: OXIDATIVE PHOSPHORYLATION
Proton gradient is harnessed by ATP synthase to make ATP

REGULATION OF ETC & OXIDATIVE PHOSPHORYLATION

§ ETC
§ Availability of ADP, inorganic P, O2 and NADH.
§ Low levels any of the compound will decrease the
formation of ATP
§ When ATP is consumed rapidly (cell active), high ADP will
activate ATP synthesis.
§ ETC dependent on ADP for ATP synthesis.

UNCOUPLING PROTEINS
§ Allow the import of H+ from intermembrane space to matrix
§ No synthesis of ATP
§ Energy released as heat to maintain body temperature in
hibernating animals, cold adapted mammals & some
newborns. (non-shivering thermogenesis)

2,4- DINITROPHENOL (DNP)

§ Uncoupled protein
§ Pills for rapid weight loss
§ Increased the metabolism of fat