Molecular evalution

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Slide 1: 

Classification of Proteases Metal Protease Serine Protease Cysteine Protease Aspartyl Protease Family Example  Mechanism Specificity Inhibitor Juang RH (2004) BCbasics

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Modification of Subtilisin and Its Activity Change No enzyme 1 Asn155 → Leu ● ● ● 10,000,000 (Asn155 stabilizes transition state) His & Asp → Ala ● ○ ○ 37,000 Ser, His & Asp → Ala ○ ○ ○ 4,000 Subtilisin ● ● ● 10,000,000,000 Active Site Relative Modification Triad: Ser His Asp activity Ser → Ala ○ ● ● 5,000 Asp → Ala ● ● ○ 330,000 Adapted from Dressler & Potter (1991) Discovering Enzymes, p.245

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Serine Protease and AchE Chymotrypsin – Gly – Asp – Ser – Gly – Gly – Pro – Leu – Trypsin – Gly – Asp – Ser – Gly – Gly – Pro – Val – Elastase – Gly – Asp – Ser – Gly – Gly – Pro – Leu – Thrombin – Gly – Asp – Ser – Gly – Gly – Pro – Phe – Plasmin – Gly – Asp – Ser – Gly – Gly – Pro – Leu – Acetylcholinesterase – Gly – Glu – Ser – Ala – Gly – Gly – Ala – Chymotrypsin – Val – Thr – Ala – Ala – His – Cys – Gly – Trypsin – Val – Ser – Ala – Gly – His – Cys – Tyr – Elastase – Leu – Thr – Ala – Ala – His – Cys – Ile – Thrombin – Leu – Thr – Ala – Ala – His – Cys – Leu – Plasmin – Leu – Thr – Ala – Ala – His – Cys – Leu – Acetylcholinesterase – – – – – – – – – – – – – His – – – – – – – – Ser 195 Chymotrypsin – Thr – Ile – Asn – Asn – Asp – Ile – Thr – Trypsin – Tyr – Leu – Asn – Asn – Asp – Ile – Met – Elastase – Ser – Lys – Gly – Asn – Asp – Ile – Ala – Thrombin – Asn – Leu – Asp – Arg – Asp – Ile – Ala – Plasmin – Phe – Thr – Arg – Lys – Asp – Ile – Ala – Acetylcholinesterase – – – – – – – – – – – – – – Asp – – – – – – – His 57 Asp 102 Adapted from Dressler & Potter (1991) Discovering Enzymes, p.244

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H AchE Has Similar Catalytic Mechanism H-O-H H2O Adapted from Dressler & Potter (1991) Discovering Enzymes, p.243 ↓Deacylation Acylation↑

Slide 5: 

Different Enzymes Might Adopt Same Mechanism Hi, Everybody! ← Useful ↙ Amusing Juang RH (2004) BCbasics

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Convergent and Divergent Trypsin Chymotrypsin Elastase Thrombin Plasmin Acetylcholinesterase Thyroglobulin Ester bond Peptide bond hydrolyze acetylcholine Serine Protease Juang RH (2004) BCbasics

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