1lzv
From Proteopedia
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==Overview== | ==Overview== | ||
| - | We have prepared a site-specific mutant of human carbonic anhydrase (HCA) | + | 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 a different isozyme, we have placed histidine residues in HCA III at positions 64 and 67 and in another mutant at positions 64 and 7. Each of these histidine residues can act as a proton transfer group in catalysis when it is the only nonliganding histidine in the active site cavity, except His(7) in HCA III. Using an (18)O exchange method to measure rate constants for intramolecular proton transfer, we have found that inserting two histidine residues into the active site cavity of either isozyme II or III of carbonic anhydrase results in rates of proton transfer to the zinc-bound hydroxide that are antagonistic or suppressive with respect to the corresponding single mutants. The crystal structure of Y7H HCA II, which contains both His(7) and His(64) within the active site cavity, shows the conformation of the side chain of His(64) moved from its position in the wild type and hydrogen-bonded through an intervening water molecule with the side chain of His(7). This suggests a cause of decreased proton transfer in catalysis. |
==Disease== | ==Disease== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: An, H.]] | [[Category: An, H.]] | ||
| - | [[Category: Duda, D | + | [[Category: Duda, D M.]] |
| - | [[Category: Laipis, P | + | [[Category: Laipis, P J.]] |
[[Category: McKenna, R.]] | [[Category: McKenna, R.]] | ||
[[Category: Pathak, Y.]] | [[Category: Pathak, Y.]] | ||
[[Category: Qian, M.]] | [[Category: Qian, M.]] | ||
| - | [[Category: Silverman, D | + | [[Category: Silverman, D N.]] |
| - | [[Category: Tu, C | + | [[Category: Tu, C K.]] |
| - | [[Category: Wadhwa, N | + | [[Category: Wadhwa, N R.]] |
[[Category: Yoshioka, C.]] | [[Category: Yoshioka, C.]] | ||
[[Category: ZN]] | [[Category: ZN]] | ||
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[[Category: zinc metalloenzyme]] | [[Category: zinc metalloenzyme]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:50:01 2008'' |
Revision as of 11:50, 21 February 2008
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Site-Specific Mutant (Tyr7 replaced with His) of Human Carbonic Anhydrase II
Contents |
Overview
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 a different isozyme, we have placed histidine residues in HCA III at positions 64 and 67 and in another mutant at positions 64 and 7. Each of these histidine residues can act as a proton transfer group in catalysis when it is the only nonliganding histidine in the active site cavity, except His(7) in HCA III. Using an (18)O exchange method to measure rate constants for intramolecular proton transfer, we have found that inserting two histidine residues into the active site cavity of either isozyme II or III of carbonic anhydrase results in rates of proton transfer to the zinc-bound hydroxide that are antagonistic or suppressive with respect to the corresponding single mutants. The crystal structure of Y7H HCA II, which contains both His(7) and His(64) within the active site cavity, shows the conformation of the side chain of His(64) moved from its position in the wild type and hydrogen-bonded through an intervening water molecule with the side chain of His(7). This suggests a cause of decreased proton transfer in catalysis.
Disease
Known disease associated with this structure: Osteopetrosis, autosomal recessive 3, with renal tubular acidosis OMIM:[611492]
About this Structure
1LZV is a Single protein structure of sequence from Homo sapiens with as ligand. Active as Carbonate dehydratase, with EC number 4.2.1.1 Full crystallographic information is available from OCA.
Reference
Kinetic analysis of multiple proton shuttles in the active site of human carbonic anhydrase., Tu C, Qian M, An H, Wadhwa NR, Duda D, Yoshioka C, Pathak Y, McKenna R, Laipis PJ, Silverman DN, J Biol Chem. 2002 Oct 11;277(41):38870-6. Epub 2002 Aug 8. PMID:12171926
Page seeded by OCA on Thu Feb 21 13:50:01 2008
