Amino Acid Sequence and Activity of Green Turtle (Chelonia Mydas) Lysozyme

Overview

Journal Protein J

Publisher Springer

Specialty Biochemistry

Date 2006 Sep 2

PMID 16947076

Citations 3

Authors

Yuki Chijiiwa

Shunsuke Kawamura

Takao Torikata

Tomohiro Araki

Affiliations

Soon will be listed here.

Abstract

Green turtle lysozyme purified from egg white was sequenced and analyzed its activity. Lysozyme was reduced and pyridylethylated or carboxymethylated to digest with trypsin, chymotrypsin and V8 protease. The peptides yielded were purified by RP-HPLC and sequenced. Every trypsin peptide was overlapped by chymotrypsin peptides and V8 protease peptides. This lysozyme is composed of 130 amino acids including an insertion of a Gly residue between 47 and 48 residues when compared with chicken lysozyme. The amino acid substitutions were found at subsites E and F. Namely Phe34, Arg45, Thr47, and Arg114 were replaced by Tyr, Tyr, Pro, and Asn, respectively. The time course using N-acetylglucosamine pentamer as a substrate showed a reduction of the rate constant of glycosidic cleavage and transglycosylation and increase of binding free energy for subsite E, which proved the contribution of amino acids mentioned above for substrate binding at subsites E and F.

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