Asp 280 residue is important in the activity of the Escherichia coli leader peptidase
Experimental & Molecular Medicine
;
: 64-69, 1999.
Article
in English
| WPRIM
| ID: wpr-56736
ABSTRACT
Leader peptidase is a novel serine protease in Escherichia coli, which catalyzes the cleavage of amino-terminal signal sequences from exported proteins. It is an integral membrane protein containing two transmembrane segments with its carboxy-terminal catalytic domain residing in the periplasmic space. Recently, the x-ray crystal structure of signal peptidase-inhibitor complex showed that Asp 280, a highly conserved consensus sequence of E. coli leader peptidase is the closest charged residue in the vicinity of two catalytic dyad, Ser 90 and Lys 145, and it is likely held in place by a salt bridge to Arg 282. Possible roles of Asp 280 and Arg 282 in the structure-catalytic function relationship were investigated by the site-directed mutagenesis of Asp 280 substituted with alanine, glutamic acid, glycine, or asparagine and of Arg 282 with methionine. All of mutants purified with nickel affinity chromatography were inactive using in vitro assay. It is surprising to find complete lose of activity by an extension of one carbon units in the mutant where Asp 280 is substituted with glutamic acid. These results suggest that Asp 280 and Arg 282 are in a sequence which constitutes catalytic crevice of leader peptidase and are essential for maintaining the conformation of catalytic pocket.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Oligonucleotides
/
Protein Precursors
/
Structure-Activity Relationship
/
Bacterial Outer Membrane Proteins
/
Serine Endopeptidases
/
Blotting, Western
/
Mutagenesis, Site-Directed
/
Aspartic Acid
/
Escherichia coli
/
Micrococcal Nuclease
Language:
English
Journal:
Experimental & Molecular Medicine
Year:
1999
Type:
Article
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