| Project/Area Number |
09672256
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | Tokyo University of Pharmacy and Life Science |
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Principal Investigator |
BEPPU Masatoshi Tokyo University of Pharmacy and Life Science, School of Pharmacy, Professor, 薬学部, 教授 (60114633)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1997: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Protease / Serine protease / Oxidative stress / ACPH / Proteolysis / Erythrocyte protease / Glycation / Oxidized protein / グリケーション |
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| Research Abstract |
In this project, we have carried out studies on the primary structure and the function of a serine protease of human erythrocytes that preferentially degrades oxidatively modified proteins (Oxidized Protein Hydrolase : OPH), and obtained the following findings.
1) STUDY ON THE PRIMARY STRUCTURE
The enzyme OPH was purified from the cytosol of human erythrocytes according the method we have previously established, and some peptide fragments were separated after degradation of the purified enzyme by endopeptidase. The amino acid sequence of these peptides were found to be highly homologous to the sequence of human acylpeptide hydrolase (ACPH), an enzyme that hydrolyzes an acylated N-terminal amino acid residue of protein. We, then, prepared recombinant human ACPH (rACPH), by cloning of human ACPH cDNA from cDNAs of human erythroblasts based on the known nucleotide sequence of human ACPH.The rACPH exhibited proteolytic activity on oxidized and glycated proteins, and reacted with specific antibody to OPH.Conversely, OPH exhibited ACPH activity. We concluded that OPH is ACPH.This is the first work that elucidated the structure of oxidized protein-selective protease, although presence of several kinds of such proteases have been reported.
2) STUDY ON ITS FUNCTION
OPH purified from human erythrocyte cytosol strongly bound to oxidized membranes of human erythrocytes. This enzyme was found to be present more on oxidized erythrocyte membranes than on unoxidized membranes. These results suggested that OPH plays an important role in degrading oxidized proteins of cell membrane.
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