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Parts of an Enzyme
Active site- portion of the enzyme where
Allosteric site
Other site besides the active site
SUBSRTATE
ENZYME
SUBSRTATE
ENZYME
INDUCED FIT
Cofactor
Nonprotein molecule which maybe necessary for
enzyme activity
Coenzyme- Organic cofactor (NAD, heme,
vitamins)
Activators- Inorganic cofactor (eg. Chloride or
Cofactor
SUBSRTATE
ENZYME
APOENZYME
HOLOENZYME
NOTE:
Holoenzyme- complete, active enzyme system Proenzyme (zymogen)- inactive enzyme
Isoenzyme- enzymes that have different physical properties but have the same catalytic functions
Classification of Enzymes
OXIDOREDUCTASES
Catalyze an oxidation-reduction reaction between
2 substrates
TRANSFERASES
Catalyze the transfer of a group from one
substrate to another
HYDROLASES
Catalyzes hydrolysis of various bonds
LYASES
Removal of groups without hydrolysis
ISOMERASES
Catalyze the interconversion of geometric, optical
or positional isomers
LIGASES
Catalyze the joining of two substrate molecules,
pH ( 7.0- 8.0)
TEMPERATURE ENZYME CONCENTRATION
COFACTORS
INHIBOTORS
Competitive inhibitors Noncompetitive inhibitor Uncompetitive inhibitor
Competitive Inhibitors
Competes with substrate for binding to active site
Non-Competitive Inhibitors
Does not bind to the active site, but removes
Uncompetitive Inhibitors
Inhibitor binds to E-S complex preventing
dissociation
Moderate
AST (Liver, heart, skeletal muscle)) CK (Heart, skeletal muscle, brain) ALP (liver, bone, kidney)
Low
LD (In ALL tissues)
ALT (Liver)
LPS (Pancreas)
AST
LD (LDH)
GGT
ChE
Pagets disease
Rickets Osteoblastic tumors Cancer Sever fracture Liver disease
Collection:
AVOID using citrate, oxalate, EDTA
ALT
Pyruvic Acid
Method:
Reitman Frankel
Addition of DNPH (2,4- dihydrophenylhydrazine) to pyruvic acid to produce color
ALT (Liver)
Substrate End Product
Location High Concentration Significance
Alanine - Keto
Aspartate keto
Pyruvic Acid
Heart, LIVER LIVER
Oxaloacetic Acid
HEART, Liver HEART Mycocardial Infarction
Liver Disease
pyruvate
clot must be separated from serum quickly to prevent increase in LD1 and LD2
Storage: 25C upto 24 hours Only LD4 and LD5 are utilized to test LIVER
-nitroanilide
Pseudocholinesterase (ChE)
Cholinesterase (true and Pseudo)
Cleaves Acetylcholine: the bodys major
Significant decrease seen after exposure to phosphorus compounds found in insecticide, nerve gases and pesticide.
Also seen in anesthetic poisoning. Substrate: acetylcholine Method:
Michael
Ellman
AST
LD (LDH) 1 & 2 LD/HBD ratio MI Biomarkers
Catalyzes
phosphocreatine + ADP
Methods:
Tanzer-Gilvarg: (forward)
creatine + ATP
Phospho Kinase + LD
Oliver- Rosalki: (reverse)
Lactate + NAD
Hexokinase + G6PD
disease
Collection: AVOID HELOLYSIS Inhibited by all anticoagulants except heparin
hemolyzed) 50% MI: flipped ratio in 48hours 80% MI: flipped ratio in 72 hours
Non-Enzymatic MI Markers
MYOGLOBIN
Major protein responsible for oxygen supply of
striated muscle
TROPONIN
the troponin complex is a component of the thin
Amylase
found in the SALIVARY GLANDS and
PANCREAS
Breaks down starch to simple sugars Substrate: starch
starch/ amylum
AMS
maltose
Maltase
glucose
Substrates:
Pancreatic AMS: diastase Salivary AMS: ptyalin
found in urine macroAMS: bound to IgG, IgA. High MW. Measured in serum
Methods
Saccharogenic
Measures amount of maltose produced (glucose: Somogyi)
Iodometric/amyloclastic
Chromogenic
Kinetic Method
Lipase
Breaksdown Triglyceride into fatty acids and
glycerol
Significance: Acute Pancreatitis Substrate: Olive Oil End Product: Fatty Acids Methods:
Cherry-Crandall Sigma-Tietz Titration
ACP (pACp)
Optimum pH: 5 Very Labile, Add 5M acetate buffer or citrate
tablet to preserve
ACP are found in Prostate, RBC, bone, liver,
kidneys, platelets
Substrate: organic Phosphate such as -glyceroPO4 and -nitrophenylPO4
Method
Chemical Inhibition Test
If Total ACP is normal: Stop test If elevated: suggestive of prostatic CA do p-ACP by Chemical Inhibition Test
Cu++ RBC-ACP
p-ACP
Inactivated (+)
Unaffected (-)
PSA
(Prostate-Specific Antigen)
proteases uniquely produced form the epithelial cells of the prostate gland
Most useful TM for Prostate Cancer Drawback: weak in distinguishing prostate CA
Good luck and God bless! Study well and Pray Harder!
-Maam Siv