PDBsum entry 1iav

Hydrolase

PDB id: 1iav

Contents

Protein chain

269 a.a. *

Ligands

SO4

Metals

_CA ×2

Waters ×111

* Residue conservation analysis

PDB id:

1iav

Name:

Hydrolase

Title:

Structure on native (asn 87) subtilisin from bacillus lentus

Structure:

Subtilisin savinase. Chain: a. Engineered: yes. Mutation: yes

Source:

Bacillus lentus. Organism_taxid: 1467. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.80Å R-factor: 0.148

Authors:

M.Knapp,R.Bott

Key ref:

T.Graycar et al. (1999). Engineered Bacillus lentus subtilisins having altered flexibility. J Mol Biol, 292, 97. PubMed id: 10493860 DOI: 10.1006/jmbi.1999.3033

Date:

23-Mar-01 Release date: 18-Apr-01

Supersedes:

1c13

Protein chain

P29600  (SUBS_LEDLE) -  Subtilisin Savinase from Lederbergia lenta

Seq: 269 a.a.

Struc: 269 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.3.4.21.62 - subtilisin.
• Reaction: Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyzes peptide amides.

DOI no: 10.1006/jmbi.1999.3033

J Mol Biol 292:97 (1999)

PubMed id: 10493860

Engineered Bacillus lentus subtilisins having altered flexibility.

T.Graycar, M.Knapp, G.Ganshaw, J.Dauberman, R.Bott.

ABSTRACT

The three-dimensional structures of engineered variants of Bacillus lentus subtilisin having increased enzymatic activity, K27R/N87S/V104Y/N123S/T274A (RSYSA) and N76D/N87S/S103A/V104I (DSAI), were determined by X-ray crystallography. In addition to identifying changes in atomic position we report a method that identifies protein segments having altered flexibility. The method utilizes a statistical analysis of variance to delineate main-chain temperature factors that represent significant departures from the overall variance between equivalent regions seen throughout the structure. This method reveals changes in main-chain mobility in both variants. Residues 125-127 have increased mobility in the RSYSA variant while residues 100-104 have decreased mobility in the DSAI variant. These segments are located at the substrate-binding site and changes in their mobility are believed to relate to the observed changes in proteolytic activity. The effect of altered crystal lattice contacts on segment flexibility becomes apparent when identical variants, determined in two crystal forms, are compared with the native enzyme.

Selected figure(s)

Figure 3.
Figure 3. Stereo diagram showing 2 F[o] -F[c]electron density map for the catalytic triad of RSYSA(I) contoured at 1s. The density for Ser125 is discontinuous in the electron density map of RSYSA(I) in contrast to the native density shown in Figure 1.

Figure 6.
Figure 6. Stereo diagram showing 2 F[o] -F[c]electron density map for the catalytic triad of DSAI. Partial density is observed for PMSF suggesting that the phenyl moiety is disordered. The side-chain of His is again oriented away from Ser221.

The above figures are reprinted by permission from Elsevier: J Mol Biol (1999, 292, 97-0) copyright 1999.

Figures were selected by an automated process.