Flashcards in Biochemistry Deck (556)
Molecule on which enzyme performs a reaction
What is lysozyme?
An enzyme found in tissue fluids/secretions, tears, saliva, nasal mucus that protects against bacteria by causing cell to lyse and lose cell content
What is peptidoglycan?
Polysaccharide formed from polymerisation of monosaccharides N-acetylglucosamine (NAG) and N-acetylmuramic acid (NAM)
Describe the structure of bacterial cell wall
Made of peptidoglycan - retains high internal osmotic pressure
Layers of repeating peptidoglycan units with peptide chain bound to NAM
Penta-glycine cross-links between last residue of chain and second last of opposite chain - added strength
Describe alpha and beta linkages
Alpha: H atoms of the C atoms in bond in cis formation
Beta: H atoms of C atoms in bond in trans formation
Describe the lysis of the cell wall
Lysozyme hydrolyses B1:4 bond (between C1 and C4) in glycan between NAG and NAM
Occurs rapidly at close to pH 7
What else will lysozyme hydrolyse?
Oligo NAG (min 6 residues)
Fit into groove round enzyme that closes on the strand
What are serine proteases?
Enzymes with hyperactive serine residue at active site and appropriately spaced histidine and aspartate resides
Name some serine proteases
How do serine proteases work?
Catalyse by breaking peptide bond using charge relay system
Describe the charge relay system
Flow of electrons between 3 AAs making serine hyperactive
AAs are far apart in primary structure but close in 3D structure due to protein folding
Why are proteolytic enzymes secreted as zymogens?
As are hazardous to body so secreted as zymogens that are activated in gut lumen
Give examples of proteolytic enzymes
Describe the activation of pepsinogen
Chief cells in gastric gland produce pepsinogen
Parietal cells produce HCl hydrolyses some peptide bonds to produce pepsin
Left over pepsinogen is hydrolysed again to produce more pepsin
Describe the activation of trypsinogen
Pancreatic duct secretes pancreatic zymogens
Some zymogens converted to trypsinogen which is converted to trypsin by enterokinase (produced by brush border)
Trypsin activates more enzymes such as chymotrypsinogen
Describe the activation of chymotrypsinogen
Inactive state: single polypeptide, 245AAs with 5 SS bridges, three portions of AS caN'T come together
Trypsin cuts bond between Arg15 and next residue - pi-chymotrypsin left
Cut at Leu13 removes dipeptide bond
Cut at Tyr146 and Asn148 removes another - a-chymotrypsin
3 parts held by SS bridges
Active site resides come together
Charge transfer complex with hyperactive serine formed
How is specificity of enzymes dictated?
Pocket close to AS into which an AA side chain may fit thus size and nature of pocket are important
Describe the specificity of trypsin, chymotrypsin and elastase
Trypsin: cuts basic R groups; Asp at bottom of pocket
Chymotrypsin: cuts hydrophobic R; hydrophobic pocket
Elastase: cuts small R; small pocket
Describe the steady state hypothesis
In enzyme catalysed reaction, enzyme and substrate form complex which is a transient state (either breaking down or reversing)
Initially conc. of complex will increase but will become steady as when broken down enzyme will form new complex
What is Vmax?
The velocity a reaction approaches as substrate conc. is increased
What is Michaelis constant (Km)?
Substrate conc. at 1/2Vmax
Dependent on substrate and enzyme
Measures affinity of enzyme for substrate - a high Km confers low affinity
What is the relationship between substrate conc. and velocity?
v = Vmax[S]/(Km+[S])
When [s]=Km v = 1/2Vmax
When [s]>>Km v ~ Vmax
What is the importance of Km?
Can use to compare 1 enzyme's affinity for different substrates or
Multiple enzymes affinities' for 1 substrate
Widespread in tissue
Low Km = high affinity
Found in liver and pancreas
High Km = low affinity only active at high glucose conc.
At high conc. glucose stored in liver as glycogen, insulin produced by pancreas
Explain the influencing factors of enzymes
Optimum temp. and pH range above optimum will denature and activity will suddenly drop
Increasing temp. increases energy of collisions, internal energy of reactants and number of collisions
What are lipids?
Diverse class of insoluble compounds that don't form polymers
Are hydrophobic (mostly hydrocarbons) and soluble in organic solvents (ether, acetone, chloroform)
Give examples of lipids
Fats, oils, fatty acids, triacylglycerols, glycolipids, phospholipids, steroids
What is the main function of fatty acids?
Catabolised generating ATP or used to synthesise triglycerides and phospholipids