Kinetic Analysis of Multiple Proton Shuttles in the Active Site of Human Carbonic Anhydrase
作者： Chingkuang Tu , Minzhang Qian , Haiqian An , Nina R. Wadhwa , David Duda
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摘要: We have prepared a site-specific mutant of human carbonic anhydrase (HCA) II with histidine residues at positions 7 and 64 in the active site cavity. Using different isozyme, we placed HCA III 67 another 7. Each these can act as proton transfer group catalysis when it is only nonliganding cavity, except His(7) III. an (18)O exchange method to measure rate constants for intramolecular transfer, found that inserting two into cavity either isozyme or results rates zinc-bound hydroxide are antagonistic suppressive respect corresponding single mutants. The crystal structure Y7H II, which contains both His(64) within shows conformation side chain moved from its position wild type hydrogen-bonded through intervening water molecule His(7). This suggests cause decreased catalysis.
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