Purification and further characterization of enteropeptidase from porcine duodenum.

Purification and further characterization of enteropeptidase from porcine duodenum. Research Abstract Details 

Research Abstract Table of Contents

Jump to the:

Purification and further characterization of enteropeptidase from porcine duodenum. Abstract Text:

M Matsushima,M Ichinose,N Yahagi,S Tsukada-Kato,K Miki,M Omata,Y T Kim,H Ito,T Takahashi,Y Sakurai,Y Tsuchiya,S B Athauda,H Inoue,K Takahashi,

Enteropeptidase [EC 3.4.21.9] is a membrane-bound serine endopeptidase present in the duodenum that converts trypsinogen to trypsin. We previously cloned the cDNA of the porcine enzyme and deduced its entire amino acid sequence [M. Matsushima et al. (1994) J. Biol. Chem. 269, 19976-19982]. In the present study, we purified the porcine enzyme approximately 2,200-fold in a 12% yield from a duodenal mucosal extract to apparent homogeneity by an improved procedure comprising four steps of chromatography including benzamidine-Sepharose affinity chromatography. Lectin blotting analysis suggested that the enzyme is glycosylated mainly with N-linked carbohydrate chains of the tri- and/or tetraantennary complex type. The H and L chains of the enzyme were separated into two major bands upon SDS-PAGE under reducing conditions, suggesting that the enzyme mainly comprises two isoforms, a higher molecular weight form and a lower molecular weight form. The enzyme was also separated by lectin affinity chromatography into two major fractions, named isoforms I and II, which corresponded to the higher and lower molecular weight forms, respectively. These two isoforms appeared to be different only in the carbohydrate moiety, having essentially the same enzymatic properties. The enzyme was optimally active at pH 8.0 toward Gly-Asp-Asp-Asp-Asp-Lys-beta-naphthylamide, and was inhibited strongly by various serine proteinase inhibitors. Furthermore, it was also strongly inhibited by E-64 [L-trans-epoxysuccinyl-leucylamide-(4-guanido)-butane], a cysteine proteinase inhibitor. Substrate specificity studies involving various synthetic peptides indicated that acidic residues at the P2, P3, and/or P4 positions are especially favorable for maximal activity, but are not absolutely necessary, at least in the cases of peptide substrates.

Purification and further characterization of enteropeptidase from porcine duodenum. Publishing Authors By Initials

M Matsushima,M Ichinose,N Yahagi,S Tsukada-Kato,K Miki,M Omata,YT Kim,H Ito,T Takahashi,Y Sakurai,Y Tsuchiya,SB Athauda,H Inoue,K Takahashi,

For similar animals: chordata: vertebrates: mammals: artiodactyla: swine research abstracts see: animals: chordata: vertebrates: mammals: artiodactyla: swine research

PUBMED ID PMID:

MEDLINE DATE:

Purification and further characterization of enteropeptidase from porcine duodenum. Journal Published:

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

Journal: Journal of biochemistry

VOLUME: 125

Page Numbers: 947-51

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: May

YEAR: 1999

Purification and further characterization of enteropeptidase from porcine duodenum. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 376600

Purification and further characterization of enteropeptidase from porcine duodenum. Keywords Mesh Terms:

KEYWORDS: Swine

MESH TERMS: isolation & purification

Chemical & Substance for Abstract: Purification and further characterization of enteropeptidase from porcine duodenum. Information

Substance Name: Enteropeptidase

Registry Number: EC 3.4.21.9

Grant and Affiliation Information for Purification and further characterization of enteropeptidase from porcine duodenum.

AFFILIATION: Department of Internal Medicine, Faculty of Medicine, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo, 192-0392, Japan.

Country: JAPAN

AGENCY:

GRANT:

ACRONYM:

MEDLINETA: J Biochem

REFSOURCE:

DATABASENAME:

ACCESSION NUMBER:

Number Hits: 0

Purification and further characterization of enteropeptidase from porcine duodenum Related Publications

• A noncanonical E-box enhancer drives mouse Period2 circadian oscillations in vivo.
• Bone morphogenetic protein 2 stimulates osteoclast differentiation and survival supported by receptor activator of nuclear factor-kappaB ligand.
• Circadian rhythms: the cancer connection.
• Comparison between stabilometry with and without head tilts in a roll plane.
• Effect of Escherichia coli wild type or its derivative with high nitrite reductase activity on in vitro ruminal methanogenesis and nitrate/nitrite reduction.
• Effect of gravity on apical dominance in Pharbitis nil.
• Effect of oral branched chain amino acid-rich nutrient administered during endoscopic injection sclerotherapy of cirrhotic patients.
• Forward genetic screens to identify circadian rhythm mutants in mice.
• Genome-wide epistatic interaction analysis reveals complex genetic determinants of circadian behavior in mice.
• Identification of the cDNA of differentially expressed genes in the transition zone of cucumber seedlings by fluorescent differential display.
• In vivo bone-forming capacity of human bone marrow-derived stromal cells is stimulated by recombinant human bone morphogenetic protein-2.
• Isolation of auxin efflux carrier cDNA from cucumber.
• Methods to record circadian rhythm wheel running activity in mice.
• Mice lacking smooth muscle calponin display increased bone formation that is associated with enhancement of bone morphogenetic protein responses.
• Mouse chimeras and their application to circadian biology.
• Novel hydrotropism mutants of Arabidopsis thaliana and their altered waving response and phototropism.
• Oral myiasis treated with ivermectin: case report.
• Osteoblasts provide a suitable microenvironment for the action of receptor activator of nuclear factor-kappaB ligand.
• Osteoprotegerin regulates bone formation through a coupling mechanism with bone resorption.
• Pax 2 expression in mesodermal segmentation and its relationship with EphA4 and Lunatic-fringe during chicken somitogenesis.
• Physiological and genetic characterization of hydrotropic mutants of Arabidopsis thaliana.
• Real-time luminescence reporting of circadian gene expression in mammals.
• Regulations: what next?
• The effect of light on the gravimorphogenesis of cucumber seedlings.
• The mouse Clock mutation reduces circadian pacemaker amplitude and enhances efficacy of resetting stimuli and phase-response curve amplitude.
• [Bone morphogenetic protein (BMP): from basic studies to clinical approaches]
• [Differential expression of a homing-related molecule repertoire among umbilical cord blood, mobilized peripheral blood and bone marrow-derived hematopoietic stem/progenitor cells.]
• [Gravitaxis of paramecium: testing the hypotheses by free-fall experiments]
• [Sequence variability in Borna disease virus ORF II from human beings]
• [The effects of space environment on differentiation and mutation frequency in Dictyostelium discoideum]