Dipeptidyl Peptidase III from Rat Liver Cytosol: Purification, Molecular Cloning and Immunohistochemical Localization
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Iwao Ohkubo
Abstract
Dipeptidyl peptidase III (DPP III) was purified to homogeneity from rat liver cytosol. The calculated molecular weight of the purified enzyme was 82845.6 according to TOF-MS and 82000 on non-denaturing PAGE, and 82000 on SDS-PAGE in the absence or presence of Β-mercaptoethanol. These findings suggest that the enzyme exists in a monomeric form in rat liver cytosol. The enzyme rapidly hydrolyzed the substrate Arg-Arg- MCA and moderately hydrolyzed Gly-Arg-MCA in the pH range of 7.5 to 9.5. The Km, kcat and kcat/Km values of DPP III at optimal pH (pH 8.5) were 290μM, 18.0 s−1 and 62.1 s−1 .nm−1 for Arg-Arg-MCA and 125μM, 4.53 s−1 and 36.2 s−1 .nm−1 for Ala-Arg-MCA, respectively. DPP III was potently inhibited by EDTA, 1,10-phenanthroline, DFP, PCMBS and NEM. These findings suggest that DPP III is an exo-type peptidase with characteristics of a metallo- and serine peptidase. For further information on the molecular structure, we screened a rat liver cDNA library using affinity-purified anti-rat DPP III rabbit IgG antibodies, determined the cDNA structure and deduced the amino acid sequence. The cDNA, designated as λRDIII-11, is composed of 2640 bp and encodes 738 amino acids in the coding region. Although the enzyme has a novel zinc-binding motif, HEXXXH, DPP III is thought to belong to family 1 in clan MA in the metalloprotease kingdom.
The DPP III antigen was detected in significant amounts in the cytosol of various rat tissues by immunohistochemical examination.
Copyright © 1999 by Walter de Gruyter GmbH & Co. KG
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- Genomic Expansion Across the Albumin Gene Family on Human Chromosome 4q Is Directional
- Comparison of the Tamoxifen Regulated Chimeric Cre Recombinases MerCreMer and CreMer
- The Human Cathepsin F Gene a Fusion Product between an Ancestral Cathepsin and Cystatin Gene
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- Molecular Cloning and Characterization of a cDNA Encoding a Transferrin Homolog from Bombyx mori
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- Acknowledgement
- Content Index
- Author Index
- Subject Index
Artikel in diesem Heft
- To Our Authors, Readers and Subscribers
- Editor's Note
- A Chimpanzee Millennium
- Molecular Genetics of Dopa-Responsive Dystonia
- In Vitro Transcription of a TATA-Less Promoter: Negative Regulation by the Not1 Protein
- Fast Control of DNA Replication in Response to Hypoxia and to Inhibited Protein Synthesis in CCRF-CEM and HeLa Cells
- The Inhibition of NF-B Activation Pathways and the Induction of Apoptosis by Dithiocarbamates in T Cells Are Blocked by the Glutathione Precursor N-Acetyl-L-Cysteine
- Xylose Utilisation: Cloning and Characterisation of the Xylose Reductase from Candida tenuis
- Xylose Utilisation: Cloning and Characterisation of the Xylitol Dehydrogenase from Galactocandida mastotermitis
- Thermodynamic Properties and DNA Binding of the ParD Protein from the Broad Host-Range Plasmid RK2/RP4 Killing System
- Dipeptidyl Peptidase III from Rat Liver Cytosol: Purification, Molecular Cloning and Immunohistochemical Localization
- Genomic Expansion Across the Albumin Gene Family on Human Chromosome 4q Is Directional
- Comparison of the Tamoxifen Regulated Chimeric Cre Recombinases MerCreMer and CreMer
- The Human Cathepsin F Gene a Fusion Product between an Ancestral Cathepsin and Cystatin Gene
- Characterization of a New Member of the TNF Family Expressed on Antigen Presenting Cells
- Vascular Endothelial Growth Factor (VEGF) and Its Receptors in Tumor-Bearing Dogs
- Molecular Cloning and Characterization of a cDNA Encoding a Transferrin Homolog from Bombyx mori
- Mitochondria-Derived and Extra-Mitochondrial Human Type-1 Porin Are Identical as Revealed by Amino Acid Sequencing and Electrophysiological Characterisation
- Acknowledgement
- Content Index
- Author Index
- Subject Index
