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PDBsum entry 1lzv

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protein metals links
Lyase PDB id
1lzv
Jmol
Contents
Protein chain
256 a.a. *
Metals
_ZN
Waters ×56
* Residue conservation analysis
PDB id:
1lzv
Name: Lyase
Title: Site-specific mutant (tyr7 replaced with his) of human carbonic anhydrase ii
Structure: Carbonic anhydrase ii. Chain: a. Synonym: carbonate dehydratase ii, ca-ii. Engineered: yes. Mutation: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 562.
Resolution:
2.30Å     R-factor:   0.186     R-free:   0.251
Authors: C.K.Tu,M.Qian,H.An,N.R.Wadhwa,D.M.Duda,C.Yoshioka,Y.Pathak, R.Mckenna,P.J.Laipis,D.N.Silverman
Key ref:
C.Tu et al. (2002). Kinetic analysis of multiple proton shuttles in the active site of human carbonic anhydrase. J Biol Chem, 277, 38870-38876. PubMed id: 12171926 DOI: 10.1074/jbc.M205791200
Date:
11-Jun-02     Release date:   23-Oct-02    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
P00918  (CAH2_HUMAN) -  Carbonic anhydrase 2
Seq:
Struc:
260 a.a.
256 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Enzyme reactions 
   Enzyme class: E.C.4.2.1.1  - Carbonate dehydratase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: H2CO3 = CO2 + H2O
H(2)CO(3)
= CO(2)
+ H(2)O
      Cofactor: Zn(2+)
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     extracellular space   11 terms 
  Biological process     angiotensin-mediated signaling pathway   22 terms 
  Biochemical function     protein binding     5 terms  

 

 
    Added reference    
 
 
DOI no: 10.1074/jbc.M205791200 J Biol Chem 277:38870-38876 (2002)
PubMed id: 12171926  
 
 
Kinetic analysis of multiple proton shuttles in the active site of human carbonic anhydrase.
C.Tu, M.Qian, H.An, N.R.Wadhwa, D.Duda, C.Yoshioka, Y.Pathak, R.McKenna, P.J.Laipis, D.N.Silverman.
 
  ABSTRACT  
 
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.
 
  Selected figure(s)  
 
Figure 1.
Fig. 1. The pH dependence of R[H2O]/[E] (s 1) catalyzed by wild type HCA II ( ), Y7H HCA II ( ), Y7H/H64A HCA II ( circle ), and H64A HCA II ( ). The values were determined by 18O exchange at 25 °C using solutions containing no buffers; ionic strength of solution was maintained at 0.2 M by addition of Na[2]SO[4]. The lines are least squares fits of Equation 6 to the data yielding the values of pK[a] of the donor and acceptor groups and the values of k[B] given in Table III. The data for H64A HCA II is from Qian et al. (30).
Figure 3.
Fig. 3. The pH dependence of R[H2O]/[E] (ms 1) catalyzed by wild type HCA III ( ), K64H HCA III ( ), R67H HCA III ( ), and the double mutant K64H/R67H HCA III ( circle ). The values were determined by 18O exchange at 25 °C using solutions containing no buffers; ionic strength of solution was maintained at 0.2 M by addition of Na[2]SO[4]. The lines are least squares fits of Equation 6 to the data yielding the values of pK[a] of the donor and acceptor groups and the values of k[B] given in Table IV.
 
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2002, 277, 38870-38876) copyright 2002.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
18175010 A.C.O'Donoghue, T.L.Amyes, and J.P.Richard (2008).
Slow proton transfer from the hydrogen-labelled carboxylic acid side chain (Glu-165) of triosephosphate isomerase to imidazole buffer in D(2)O.
  Org Biomol Chem, 6, 391-396.  
18335973 V.M.Krishnamurthy, G.K.Kaufman, A.R.Urbach, I.Gitlin, K.L.Gudiksen, D.B.Weibel, and G.M.Whitesides (2008).
Carbonic anhydrase as a model for biophysical and physical-organic studies of proteins and protein-ligand binding.
  Chem Rev, 108, 946.  
14567703 P.J.O'Brien, and T.Ellenberger (2003).
Human alkyladenine DNA glycosylase uses acid-base catalysis for selective excision of damaged purines.
  Biochemistry, 42, 12418-12429.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.

 

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