PDBsum entry 5gch

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protein Protein-protein interface(s) links
Hydrolase (serine proteinase) PDB id
Protein chains
11 a.a.
131 a.a. *
95 a.a. *
Waters ×78
* Residue conservation analysis
PDB id:
Name: Hydrolase (serine proteinase)
Title: Chemistry of caged enzymes II . Photoactivation of inhibite chymotrypsin
Structure: Gamma-chymotrypsin a. Chain: e. Gamma-chymotrypsin a. Chain: f. Gamma-chymotrypsin a. Chain: g. Ec:
Source: Bos taurus. Cattle. Organism_taxid: 9913. Organism_taxid: 9913
Biol. unit: Trimer (from PQS)
2.70Å     R-factor:   0.156    
Authors: B.L.Stoddard,D.Ringe,G.A.Petsko
Key ref:
B.L.Stoddard et al. (1990). Photolysis and deacylation of inhibited chymotrypsin. Biochemistry, 29, 8042-8051. PubMed id: 2261462 DOI: 10.1021/bi00487a008
25-Sep-89     Release date:   15-Oct-90    
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Protein chain
Pfam   ArchSchema ?
P00766  (CTRA_BOVIN) -  Chymotrypsinogen A
245 a.a.
11 a.a.
Protein chain
Pfam   ArchSchema ?
P00766  (CTRA_BOVIN) -  Chymotrypsinogen A
245 a.a.
131 a.a.
Protein chain
Pfam   ArchSchema ?
P00766  (CTRA_BOVIN) -  Chymotrypsinogen A
245 a.a.
95 a.a.
Key:    PfamA domain  Secondary structure  CATH domain

 Enzyme reactions 
   Enzyme class: Chains E, F, G: E.C.  - Chymotrypsin.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Preferential cleavage: Tyr-|-Xaa, Trp-|-Xaa, Phe-|-Xaa, Leu-|-Xaa.
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     proteolysis   1 term 
  Biochemical function     catalytic activity     2 terms  


DOI no: 10.1021/bi00487a008 Biochemistry 29:8042-8051 (1990)
PubMed id: 2261462  
Photolysis and deacylation of inhibited chymotrypsin.
B.L.Stoddard, J.Bruhnke, P.Koenigs, N.Porter, D.Ringe, G.A.Petsko.
Inhibited chymotrypsin was reactivated through the photolysis of the covalently bound light-reversible cinnamates described in our previous paper [Stoddard, B.L., Bruhnke, J., Porter, N.A., Ringe, D., & Petsko, G. (1990) Biochemistry 29, 4871-4879]. The light-induced deacylation was accomplished both in solution and in protein crystals, with the release of inhibitor from the crystal monitored and confirmed by X-ray diffraction. The product of photolysis has been characterized as a 3-methylcoumarin, leading to a mechanism for light-driven deacylation of an internal lactonization that is dependent on the presence of an internal hydroxyl nucleophile. The acyl enzyme formed from cinnamate A is not suitable for photochemical studies, as the complex has a short half-life in solution and does not have a chromophore that is well separated from protein absorbance. Cinnamate B, with a p-diethylamino substituent, shows an enzyme deacylation rate enhancement of 10(9) for the cis photoisomer relative to the trans starting material. The half-life and deacylation rate of this compound in the E-I complex after photon absorption have been directly measured by subsecond UV absorption studies. X-ray diffraction studies of photoactivation using a flow cell show that the cinnamate B acyl enzyme complex is fully capable of light-induced isomerization and regeneration of native enzyme in the crystalline state. The E-I complex formed upon binding of cinnamate A, however, shows little if any effect from irradiation due to competitive absorbance by the highly concentrated protein at the shorter UV wavelengths. Photolysis of cinnamate B appears to occur on a time scale fast enough for applications in crystallographic studies of enzymatic intermediate-state structures.

Literature references that cite this PDB file's key reference

  PubMed id Reference
15498869 M.J.Theisen, I.Misra, D.Saadat, N.Campobasso, H.M.Miziorko, and D.H.Harrison (2004).
3-hydroxy-3-methylglutaryl-CoA synthase intermediate complex observed in "real-time".
  Proc Natl Acad Sci U S A, 101, 16442-16447.
PDB codes: 1xpk 1xpl 1xpm
12203645 K.Zou, W.T.Miller, R.S.Givens, and H.Bayley (2001).
Caged Thiophosphotyrosine Peptides.
  Angew Chem Int Ed Engl, 40, 3049-3051.  
9134755 N.A.Porter, K.A.Bush, and K.S.Kinter (1997).
Photo-reversible binding of thrombin to avidin by means of a photolabile inhibitor.
  J Photochem Photobiol B, 38, 61-69.  
8888067 B.L.Stoddard (1996).
Intermediate trapping and laue X-ray diffraction: potential for enzyme mechanism, dynamics, and inhibitor screening.
  Pharmacol Ther, 70, 215-256.  
8735386 S.D.Varfolomeyev, N.F.Kazanskaya, and N.L.Eremeev (1996).
Enzyme activity regulation by an outer physical signal.
  Biosystems, 39, 35-42.  
  1303743 L.N.Johnson (1992).
Time-resolved protein crystallography.
  Protein Sci, 1, 1237-1243.  
2062832 B.L.Stoddard, P.Koenigs, N.Porter, K.Petratos, G.A.Petsko, and D.Ringe (1991).
Observation of the light-triggered binding of pyrone to chymotrypsin by Laue x-ray crystallography.
  Proc Natl Acad Sci U S A, 88, 5503-5507.  
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