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

Journal of Biochemistry
T NonakaK Takio

Abstract

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-ornithi...Continue Reading

Citations

Apr 8, 2011·Journal of the American Society for Mass Spectrometry·Mathieu DupréChristine Enjalbal
Nov 19, 2011·Journal of the American Society for Mass Spectrometry·Mathieu DupréChristine Enjalbal
Mar 9, 2010·Journal of the American Society for Mass Spectrometry·Valerie J CarabettaIleana M Cristea
Sep 21, 2010·Journal of the American Society for Mass Spectrometry·Marco L HennrichAlbert J R Heck
Feb 7, 2009·Journal of Proteome Research·Laura HohmannDaniel B Martin
Oct 19, 2010·Chemical Communications : Chem Comm·Reinout RaijmakersAlbert J R Heck
Apr 17, 2001·Bioscience, Biotechnology, and Biochemistry·T Nishiwaki, K Hayashi
Dec 27, 2002·The Journal of General and Applied Microbiology·Tamio SaitoKoji Takio
Feb 9, 2012·Journal of Amino Acids·Kayoko M FukasawaJunzo Hirose
May 19, 2009·Journal of Bioscience and Bioengineering·Toshikazu NishiwakiTakuji Ohyama
Dec 26, 2006·Analytical Biochemistry·Gaëlle CoussotRyuji Kobayashi
Jun 9, 2009·Journal of Mass Spectrometry : JMS·Paul J BoersemaAlbert J R Heck
Apr 22, 2008·Nature Methods·Nadia TaouatasShabaz Mohammed
Oct 12, 2010·The Analyst·Chihiro NakajimaSusumu Tsunasawa
Jun 15, 2010·Journal of Proteome Research·Nadia TaouatasShabaz Mohammed
Apr 13, 2005·Journal of Proteome Research·K C Sekhar RaoMasaru Miyagi

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.