spacer
spacer

PDBsum entry 1s02
Go to PDB code: 
protein ligands metals links
Hydrolase (serine proteinase) PDB id
1s02

 

 

 

 

Loading ...

 
JSmol PyMol  
Contents
Protein chain
275 a.a. *
Ligands
SO4
Metals
_CA ×2
Waters ×118
* Residue conservation analysis
PDB id:
1s02
Name: Hydrolase (serine proteinase)
Title: Effects of engineered salt bridges on the stability of subtilisin bpn'
Structure: Subtilisin bpn'. Chain: a. Engineered: yes
Source: Bacillus amyloliquefaciens. Organism_taxid: 1390
Resolution:
1.90Å     R-factor:   0.169    
Authors: C.R.Erwin,B.L.Barnett,J.D.Oliver,J.F.Sullivan
Key ref: C.R.Erwin et al. (1990). Effects of engineered salt bridges on the stability of subtilisin BPN'. Protein Eng, 4, 87-97. PubMed id: 2127106
Date:
20-Feb-91     Release date:   15-Jan-92    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
P00782  (SUBT_BACAM) -  Subtilisin BPN' from Bacillus amyloliquefaciens
Seq:
Struc: 		382 a.a.
275 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 2 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.3.4.21.62  - subtilisin.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyzes peptide amides.

 

 
Protein Eng 4:87-97 (1990)
PubMed id: 2127106  
 
 
Effects of engineered salt bridges on the stability of subtilisin BPN'.
C.R.Erwin, B.L.Barnett, J.D.Oliver, J.F.Sullivan.
 
  ABSTRACT  
 
Variants designed using PROTEUS have been produced in an attempt to engineer stabilizing salt bridges into subtilisin BPN'. All the mutants constructed by site-directed mutagenesis were secreted by Bacillus subtilis, except L75K. Q19E, expressed as a single variant and also in a double variant, Q19E/Q271E, appears to form a stabilizing salt bridge based on X-ray crystal structure determination and differential scanning calorimeter measurements. Although the double mutant was found to be less thermodynamically stable than the wild-type, it did exhibit an autolytic stability about two-fold greater under hydrophobic conditions. Four variants, A98K, S89E, V26R and L235R, were found to be nearly identical to wild-type in thermal stability, indicative of stable structures without evidence of salt bridge formation. Variants Q271E, V51K and T164R led to structures that resulted in varying degrees of thermodynamic and autolytic instability. A computer-modeling analysis of the PROTEUS predictions reveals that the low percentage of salt bridge formation is probably due to an overly simplistic electrostatic model, which does not account for the geometry of the pairwise interactions.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
17921497 C.Deutsch, and B.Krishnamoorthy (2007).
Four-body scoring function for mutagenesis.
  Bioinformatics, 23, 3009-3015.  
11015217 K.Takano, K.Tsuchimori, Y.Yamagata, and K.Yutani (2000).
Contribution of salt bridges near the surface of a protein to the conformational stability.
  Biochemistry, 39, 12375-12381.
PDB codes: 1eq4 1eq5 1eqe
  10880975 N.Bonander, J.Leckner, H.Guo, B.G.Karlsson, and L.Sjölin (2000).
Crystal structure of the disulfide bond-deficient azurin mutant C3A/C26A: how important is the S-S bond for folding and stability?
  Eur J Biochem, 267, 4511-4519.
PDB code: 1ezl
11150607 P.N.Bryan (2000).
Protein engineering of subtilisin.
  Biochim Biophys Acta, 1543, 203-222.  
9346299 O.R.Veltman, G.Vriend, F.Hardy, J.Mansfeld, B.van den Burg, G.Venema, and V.G.Eijsink (1997).
Mutational analysis of a surface area that is critical for the thermal stability of thermolysin-like proteases.
  Eur J Biochem, 248, 433-440.  
9162945 D.Gandini, L.Gogioso, M.Bolognesi, and D.Bordo (1996).
Patterns in ionizable side chain interactions in protein structures.
  Proteins, 24, 439-449.  
8783394 H.Nakamura (1996).
Roles of electrostatic interaction in proteins.
  Q Rev Biophys, 29, 1.  
  9213423 R.S.Hodges, and R.S.Hodges (1996).
Boehringer Mannheim award lecture 1995. La conference Boehringer Mannheim 1995. De novo design of alpha-helical proteins: basic research to medical applications.
  Biochem Cell Biol, 74, 133-154.  
7749923 B.W.Matthews (1995).
Can proteins be turned inside-out?
  Nat Struct Biol, 2, 85-86.  
  8003958 Z.S.Hendsch, and B.Tidor (1994).
Do salt bridges stabilize proteins? A continuum electrostatic analysis.
  Protein Sci, 3, 211-226.  
8229092 G.Vriend, and V.Eijsink (1993).
Prediction and analysis of structure, stability and unfolding of thermolysin-like proteases.
  J Comput Aided Mol Des, 7, 367-396.  
1618293 S.Janecek, and S.Baláz (1992).
alpha-Amylases and approaches leading to their enhanced stability.
  FEBS Lett, 304, 1-3.  
1367681 A.C.Storer (1991).
Engineering of proteases and protease inhibition.
  Curr Opin Biotechnol, 2, 606-613.  
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.

 

spacer
spacer