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The pK values of 2 essential catalytic residues in RNAse are 5.4 to 6.4. Which corresponds to His12 and which to His 119
(see figure 11-10). Draw the tritration curve for these two residues.
Explain why lysozyme cleaves the artificial substrate (NAG)~4000 times more slowly than it cleaves (NAG)
Lysozyme cleaves between the 4th and 5th sugar and will
preferably bind (NAG) and distort the conformation of the
4th sugar (D) accelerates the reaction.
The lysozyme active site is arranged to cleave oligosaccharides between the 4th and 5th residues. Moreover, since the
lysozyme active site can bind at least six monosaccharide units, (NAG) would be more tightly bound to the enzyme than
(NAG) and this additional binding free energy would be applied to distorting the D ring to its half-chair conformation,
thereby facilitating the reaction
Lysozyme residues Asp 101 and Arg 114 are requried for efficient catalysis, althought they are located at some distance
from the active site GLue 35 and Asp 52. Substituting Ala for either Asp 101 or Arg 114 does not significantly alter the
enzymes tertiary structure, but it significantly reduces its catalytic activity. Explain
Asp 101 and Arg 114 form hydrogen bonds with the substrate molecule (fig 11-19). Ala cannot form theses hydrogen bonds
so the substituted enzyme is less active
Diagram the hydrogen-bonding interactions of the catalytic triad His-Lys-Ser during catalysis in a hypothetical hydrolytic
enzyme
Predict the effect of mutating Asp 102 of trypsin to Asn on substrate binding.
Little or no effect