Sandbox Reserved 1677: Difference between revisions
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== Important amino acids== | == Important amino acids== | ||
<scene name='87/873239/4_catalytic_residue/2'>There are four catalytic</scene> amino acids in AldC. | <scene name='87/873239/4_catalytic_residue/2'>There are four catalytic</scene> amino acids in AldC. | ||
''''''== Asn 159, Glu 257, Gly 288, Cys 291''' | ''''''== Asn 159, Glu 257, Gly 288, Cys 291''' | ||
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'''== Octanol Lingand (binding site) | '''== Octanol Lingand (binding site) | ||
'''Trp 160 Tyr 163, Trp 450, Phe 456, Tyr 458, met 114, leu 118 =='''''' | '''Trp 160 Tyr 163, Trp 450, Phe 456, Tyr 458, met 114, leu 118 =='''''' | ||
Apolar interactions dominate the octanal binding in the hydrophobic substrate binding pocket. A cluster of aromatic residues and two nonpolar residues (Methionine and Leucine) peovides hydophobic environment that accommodates octanal and other aliphatic aldehydes. The substrate binding site forms an aromatic box for adaptable apolar ligand interaction. | Apolar interactions dominate the octanal binding in the hydrophobic substrate binding pocket. A cluster of aromatic residues and two nonpolar residues (Methionine and Leucine) peovides hydophobic environment that accommodates octanal and other aliphatic aldehydes. The substrate binding site forms an aromatic box for adaptable apolar ligand interaction. | ||
To examine the contribution of the active site residues, a series of site directed mutants targeting residues in the NAD(H) binding site and the octanal binding site were generated. All the 31 mutants were expressed in E.coli and purified using nickel-affinity and size exclusion chromatographies. The enzyme activity screening showed that mutation of the catalytic residues in the NAD(H) binding site and octanal binding site resulted in enzyme with less than 1% of WT specific activity. | |||
https://proteopedia.org/wiki/images/4/48/Screen_Shot_2021-04-18_at_4.03.57_PM.png | https://proteopedia.org/wiki/images/4/48/Screen_Shot_2021-04-18_at_4.03.57_PM.png | ||
