Crystal structure of GyrA intein from Mycobacterium xenopi reveals structural basis of protein splicing

Details

Serval ID
serval:BIB_C859FD51D4B3
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Crystal structure of GyrA intein from Mycobacterium xenopi reveals structural basis of protein splicing
Journal
Nature Structural Biology
Author(s)
Klabunde  T., Sharma  S., Telenti  A., Jacobs, W. R., Jr. , Sacchettini  J. C.
ISSN
1072-8368 (Print)
Publication state
Published
Issued date
01/1998
Volume
5
Number
1
Pages
31-6
Notes
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S. --- Old month value: Jan
Abstract
Several genes from prokaryotes and lower eukaryotes have been found to contain an in-frame open reading frame, which encodes for an internal protein (intein). Post-translationally, the internal polypeptide auto-splices and ligates the external sequences to yield a functional external protein (extein) and an intein. Most, but not all inteins, contain, apart from a splicing domain, a separate endonucleolytic domain that enables them to maintain their presence by a homing mechanism. We report here the crystal structure of an intein found in the gyrase A subunit from Mycobacterium xenopi at 2.2 A resolution. The structure contains an unusual beta-fold with the catalytic splice junctions at the ends of two adjacent antiparallel beta-strands. The arrangement of the active site residues Ser 1, Thr 72, His 75, His 197, and Asn 198 is consistent with a four-step mechanism for the cleavage-ligation reaction. Using site-directed mutagenesis, the N-terminal cysteine, proposed as the nucleophile in the first step of the splicing reaction, was changed to a Ser 1 and Ala 0, thus capturing the intein in a pre-spliced state.
Keywords
Amino Acid Sequence DNA Gyrase DNA Topoisomerases, Type II/*metabolism/ultrastructure Deoxyribonucleases, Type II Site-Specific/metabolism/ultrastructure *Drosophila Proteins Endonucleases/chemistry/metabolism Insect Proteins/chemistry Molecular Sequence Data Mycobacterium xenopi Protein Precursors/metabolism *Protein Splicing Protein Structure, Tertiary Saccharomyces cerevisiae
Pubmed
Web of science
Create date
25/01/2008 15:45
Last modification date
20/08/2019 16:43
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