Kinetic characterization of lysine-specific metalloendopeptidases from Grifola frondosa and Pleurotus ostreatus fruiting bodies.

Kinetic characterization of lysine-specific metalloendopeptidases from Grifola frondosa and Pleurotus ostreatus fruiting bodies. Research Abstract Details 

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Kinetic characterization of lysine-specific metalloendopeptidases from Grifola frondosa and Pleurotus ostreatus fruiting bodies. Abstract Text:

T Nonaka,Y Hashimoto,K Takio,

Two zinc-metalloendopeptidases, GFMEP (accession number P81054) and POMEP (accession number P81055), from the fruiting bodies of two edible mushrooms, Grifola frondosa and Pleurotus ostreatus, respectively, specifically hydrolyze peptidyl-lysine bonds (-X-Lys-) in polypeptides. To understand detailed substrate specificities and kinetic characters of these enzymes, we have synthesized various intramolecularly quenched fluorescent peptide substrates and determined their kinetic constants with these substrates. Each synthesized fluorogenic peptide has a fluorescent residue, tryptophan, at its carboxyl terminus and a quenching group, dinitrophenyl (Dnp), at its amino terminus. Quenching of the Trp fluorescence in an intact substrate is relieved on hydrolysis of the -X-Lys- bond, giving rise to a continuous increase in fluorescence. The octapeptide substrate, Dnp-Ser-Thr-Ala-Thr-Lys-Leu-Ser-Trp, was an efficient substrate for both enzymes, the kcat/Km values being 9.8 x 10(6) and 7.0 x 10(5) M-1.s-1 for GF- and POMEP, respectively. Peptides with aspartic acid adjacent to the Lys residue were found to be poor substrates for both enzymes. Neither the shortest peptide, Dnp-Thr-Lys-Trp, nor peptides with substitution of L-Arg, L-ornithine, or D-Lys for Lys were hydrolyzed by either enzyme. These results confirmed the strict specificities of GF- and POMEP toward the peptide bond, -X-Lys-. Substitution of Co2+ for Zn2+ enhanced the activity, while the Km values were comparable. All peptides not hydrolyzed by either enzyme had inhibitory effects on GFMEP activity. The active site structure is discussed in relation to these observations.

Kinetic characterization of lysine-specific metalloendopeptidases from Grifola frondosa and Pleurotus ostreatus fruiting bodies. Publishing Authors By Initials

T Nonaka,Y Hashimoto,K Takio,

For similar biochemical phenomena, metabolism, and nutrition: biochemical phenomena: substrate specificity research abstracts see: biochemical phenomena, metabolism, and nutrition: biochemical phenomena: substrate specificity research

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Kinetic characterization of lysine-specific metalloendopeptidases from Grifola frondosa and Pleurotus ostreatus fruiting bodies. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biochemistry

VOLUME: 124

Page Numbers: 157-62

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Jul

YEAR: 1998

Kinetic characterization of lysine-specific metalloendopeptidases from Grifola frondosa and Pleurotus ostreatus fruiting bodies. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Kinetic characterization of lysine-specific metalloendopeptidases from Grifola frondosa and Pleurotus ostreatus fruiting bodies. Keywords Mesh Terms:

KEYWORDS: Substrate Specificity

MESH TERMS: metabolism

Chemical & Substance for Abstract: Kinetic characterization of lysine-specific metalloendopeptidases from Grifola frondosa and Pleurotus ostreatus fruiting bodies. Information

Substance Name: Metalloendopeptidases

Registry Number: EC 3.4.24.-

Grant and Affiliation Information for Kinetic characterization of lysine-specific metalloendopeptidases from Grifola frondosa and Pleurotus ostreatus fruiting bodies.

AFFILIATION: Department of Biochemistry, Faculty of Science, Saitama University, Urawa, Saitama, 338-8570, Japan.

Country: JAPAN

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MEDLINETA: J Biochem

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