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PDBsum entry 3tgj

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protein ligands metals Protein-protein interface(s) links
Complex (serine protease/inhibitor) PDB id
3tgj
Jmol
Contents
Protein chains
215 a.a. *
56 a.a. *
Ligands
SO4 ×2
Metals
_CA
Waters ×106
* Residue conservation analysis
PDB id:
3tgj
Name: Complex (serine protease/inhibitor)
Title: S195a trypsinogen complexed with bovine pancreatic trypsin inhibitor (bpti)
Structure: Trypsin. Chain: e. Engineered: yes. Mutation: yes. Bovine pancreatic trypsin inhibitor. Chain: i. Synonym: bpti
Source: Rattus norvegicus. Norway rat. Organism_taxid: 10116. Organ: pancreatic. Expressed in: saccharomyces cerevisiae. Expression_system_taxid: 4932. Bos taurus. Cattle. Organism_taxid: 9913
Biol. unit: Tetramer (from PQS)
Resolution:
2.20Å     R-factor:   0.197     R-free:   0.240
Authors: A.Pasternak,D.Ringe,L.Hedstrom
Key ref: A.Pasternak et al. (1999). Comparison of anionic and cationic trypsinogens: the anionic activation domain is more flexible in solution and differs in its mode of BPTI binding in the crystal structure. Protein Sci, 8, 253-258. PubMed id: 10210204 DOI: 10.1110/ps.8.1.253
Date:
16-Jul-98     Release date:   23-Dec-98    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
P00763  (TRY2_RAT) -  Anionic trypsin-2
Seq:
Struc:
246 a.a.
215 a.a.*
Protein chain
Pfam   ArchSchema ?
P00974  (BPT1_BOVIN) -  Pancreatic trypsin inhibitor
Seq:
Struc:
100 a.a.
56 a.a.
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Enzyme reactions 
   Enzyme class: Chain E: E.C.3.4.21.4  - Trypsin.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Preferential cleavage: Arg-|-Xaa, Lys-|-Xaa.
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     proteolysis   1 term 
  Biochemical function     catalytic activity     3 terms  

 

 
DOI no: 10.1110/ps.8.1.253 Protein Sci 8:253-258 (1999)
PubMed id: 10210204  
 
 
Comparison of anionic and cationic trypsinogens: the anionic activation domain is more flexible in solution and differs in its mode of BPTI binding in the crystal structure.
A.Pasternak, D.Ringe, L.Hedstrom.
 
  ABSTRACT  
 
Unlike bovine cationic trypsin, rat anionic trypsin retains activity at high pH. This alkaline stability has been attributed to stabilization of the salt bridge between the N-terminal Ile16 and Asp194 by the surface negative charge (Soman K, Yang A-S, Honig B, Fletterick R., 1989, Biochemistry 28:9918-9926). The formation of this salt bridge controls the conformation of the activation domain in trypsin. In this work we probe the structure of rat trypsinogen to determine the effects of the surface negative charge on the activation domain in the absence of the Ile16-Asp194 salt bridge. We determined the crystal structures of the rat trypsin-BPTI complex and the rat trypsinogen-BPTI complex at 1.8 and 2.2 A, respectively. The BPTI complex of rat trypsinogen resembles that of rat trypsin. Surprisingly, the side chain of Ile16 is found in a similar position in both the rat trypsin and trypsinogen complexes, although it is not the N-terminal residue and cannot form the salt bridge in trypsinogen. The resulting position of the activation peptide alters the conformation of the adjacent autolysis loop (residues 142-153). While bovine trypsinogen and trypsin have similar CD spectra, the CD spectrum of rat trypsinogen has only 60% of the intensity of rat trypsin. This lower intensity most likely results from increased flexibility around two conserved tryptophans, which are adjacent to the activation domain. The NMR spectrum of rat trypsinogen contains high field methyl signals as observed in bovine trypsinogen. It is concluded that the activation domain of rat trypsinogen is more flexible than that of bovine trypsinogen, but does not extend further into the protein core.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
19549826 E.Zakharova, M.P.Horvath, and D.P.Goldenberg (2009).
Structure of a serine protease poised to resynthesize a peptide bond.
  Proc Natl Acad Sci U S A, 106, 11034-11039.
PDB codes: 3fp6 3fp7 3fp8
18404270 A.Muhlia-Almazán, A.Sánchez-Paz, and F.L.García-Carreño (2008).
Invertebrate trypsins: a review.
  J Comp Physiol [B], 178, 655-672.  
17400917 A.E.Aleshin, S.A.Shiryaev, A.Y.Strongin, and R.C.Liddington (2007).
Structural evidence for regulation and specificity of flaviviral proteases and evolution of the Flaviviridae fold.
  Protein Sci, 16, 795-806.
PDB codes: 2ggv 2ijo
17511867 S.Q.Liu, Z.H.Meng, J.K.Yang, Y.X.Fu, and K.Q.Zhang (2007).
Characterizing structural features of cuticle-degrading proteases from fungi by molecular modeling.
  BMC Struct Biol, 7, 33.  
16542853 O.Király, L.Guan, E.Szepessy, M.Tóth, Z.Kukor, and M.Sahin-Tóth (2006).
Expression of human cationic trypsinogen with an authentic N terminus using intein-mediated splicing in aminopeptidase P deficient Escherichia coli.
  Protein Expr Purif, 48, 104-111.  
15670163 J.Arnórsdóttir, M.M.Kristjánsson, and R.Ficner (2005).
Crystal structure of a subtilisin-like serine proteinase from a psychrotrophic Vibrio species reveals structural aspects of cold adaptation.
  FEBS J, 272, 832-845.
PDB codes: 1s2n 1sh7
16038610 T.S.Zamolodchikova, E.V.Smirnova, A.N.Andrianov, I.V.Kashparov, O.D.Kotsareva, E.A.Sokolova, K.B.Ignatov, and A.D.Pemberton (2005).
Cloning and molecular modeling of duodenase with respect to evolution of substrate specificity within mammalian serine proteases that have lost a conserved active-site disulfide bond.
  Biochemistry (Mosc), 70, 672-684.  
11420435 A.Pasternak, A.White, C.J.Jeffery, N.Medina, M.Cahoon, D.Ringe, and L.Hedstrom (2001).
The energetic cost of induced fit catalysis: Crystal structures of trypsinogen mutants with enhanced activity and inhibitor affinity.
  Protein Sci, 10, 1331-1342.
PDB codes: 1f5r 1f7z 1fy8 3tgk
11685246 S.Ye, A.L.Cech, R.Belmares, R.C.Bergstrom, Y.Tong, D.R.Corey, M.R.Kanost, and E.J.Goldsmith (2001).
The structure of a Michaelis serpin-protease complex.
  Nat Struct Biol, 8, 979-983.
PDB codes: 1i99 1k9o
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. Where a reference describes a PDB structure, the PDB codes are shown on the right.