Cleavage of various peptides with pitrilysin from Escherichia coli: kinetic analyses using beta-endorphin and its derivatives.

Content Provider	Semantic Scholar
Author	Cornista, Joel C. Ikeuchi, Satoshi Haruki, Mitsuru Kohara, Atsuko Takano, Kazufumi Morikawa, Masaaki Kanaya, Shigenori
Copyright Year	2004
Abstract	Pitrilysin from Escherichia coli was overproduced, purified, and analyzed for enzymatic activity using 14 peptides as a substrate. Pitrilysin cleaved all the peptides, except for two of the smallest, at a limited number of sites, but showed little amino acid specificity. It cleaved beta-endorphin (beta-EP) most effectively, with a K(m) value of 0.36 microM and a k(cat) value of 750 min(-1). beta-EP consists of 31 residues and was predominantly cleaved by the enzyme at Lys(19)-Asn(20). Kinetic analyses using a series of beta-EP derivatives with N and/or C-terminal truncations and with amino acid substitutions revealed that three hydrophobic residues (Leu(14), Val(15), and Leu(17)) and the region 22-26 in beta-EP are responsible for high-affinity recognition by the enzyme. These two regions are located on the N- and C-terminal sides of the cleavage site in beta-EP, suggesting that the substrate binding pocket of pitrilysin spans its catalytic site.
Starting Page	800
Ending Page	803
Page Count	4
File Format	PDF HTM / HTML
Alternate Webpage(s)	http://noah.ees.hokudai.ac.jp/emb/morikawalab/pdf/BBB2004,68,10.pdf
Alternate Webpage(s)	http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.319.297&rep=rep1&type=pdf
PubMed reference number	15502359v1
Volume Number	68
Issue Number	10
Journal	Bioscience, biotechnology, and biochemistry
Language	English
Access Restriction	Open
Subject Keyword	Amino Acid Substitution Amino Acids Catalytic Domain Cleaved cell Endorphins Kinetics POMC gene beta-Endorphin pitrilysin procollagen Type I N-terminal peptide
Content Type	Text
Resource Type	Article