Enzyme kinetics：.pptVIP

* * * Specific groups aid catalysis General acid-base catalysis Covalent Catalysis Metal ion catalysis Acid-base catalysis Amino acids in acid-base catalysis Specific groups aid catalysis General acid-base catalysis Covalent Catalysis Transient covalent enzyme-substrate bond Metal ion catalysis Ionic bonds Orient substrate enzyme Stabilize charge interaction Enzyme kinetics Enzyme kinetics E+S ES E+P k1 k-1 k2 Michaelis-Menten variables Vo = initial rate of reaction [S] = substrate concentration Vmax = maximum velocity Km = constant Michaelis-Menten equation Vo= Vmax[S] Km + [S] E + S ES E + P k1 k2 k-1 Lineweaver-Burk plot Michaelis-Menten limitations Provide information about 1 substrate enzyme catalyzes reactions Not all enzymes follow Michaelis-Menten enzyme kinetics Vmax and Km do not provide information on: Number of reaction steps Rates of each reaction step Chemical nature of the reaction steps Reaction rate is related to substrate concentration At low substrate concentrations Vo= Vmax Km [S] At high substrate concentrations Vo=Vmax When [S]=Km then Vo= Vmax 1 2 Physiological consequences of Km CH3CH2OH + NAD+ CH3CHO + NADH + H+ alcohol dehydrogenase CH3CHO + NAD+ + H2O CH3COO- + NADH + 2H+ 2 forms acetaldehyde dehydrogenase low Km mitochondrial form high Km cytosolic form aldehyde dehydrogenase Significance of Km and Vmax Km Concentration of substrate when ? active sites filled Related to rate constants for individual steps Km=(k-1+ k2)/k1 Vmax Vmax = k2[Et] when k2 is rate limiting step Turnover number, kcat (sec-1) Number of substrate molecules converted to product per enzyme per second Kcat=Vmax/[Et] Catalytic efficiency (kcat/km) * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * *