| Project/Area Number |
62560070
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用生物化学・栄養化学
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| Research Institution | Ibaraki University |
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Principal Investigator |
SUGAWARA Kiyoshi Faculty of Agriculture, Ibaraki University, Professor, 農学部, 教授 (40007662)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHARA Hidenari Faculty of Agriculture, Ibaraki University, Assoc. Professor, 農学部, 助教授 (30122063)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1988: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1987: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Modification of arginine residue / Citrulline residue / HMG1 / HMG2 / HMG2 / シトルリンの定量 / HMGの修飾 |
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| Research Abstract |
1. Reliable and sensitive amino acid analyses containing citrulline; A improved method is based on a gaseous acid hydrolysis of proteins followed by the formation of PTC derivatives of the amino acids. The PTC-amino acids were subjected to HPLC separation on an ODS revers phase column followed by UV detection at 254 nm. This integrated procedure results in a rapid(20 min) analysis with a detection limit of one picomol for each amino acids containing citrulline and ornithine.
2. Detection and determination of peptidylarginine deiminase(PADase) sensitive nonhistone chromosomal proteins; PADase from rabbit skeletal muscle was immobilized on sepharose 4B and packed to small column. Nonhistone chromosomal proteins from calf thymus were run through the column at 4゜C and the eluents were fractionated to LMG and HMG proteins. The rate of midification of arginine residues of these proteins were about 11%. When HMG 1 and HMG 2 isolated from 0.75M HC10_4 extract of pig thymus were trated with PADase at 37゜C for 2hr, the rates of the modification were 33% and 44% respectively. These results indicate a part of arginine residues of HMG 1 and HMG 2 are PAGase sensitive.
3. The effect of deiminated HMG 1 and HMG 2 on the superhelical structure of DNA; This was examined using a reluxed plasmid pBR322 DNA on the basis of their electrophoretic mobility on gel. The deiminated HMG land HMG 2 showed lower functional activity towards the structure of DNA than that of the native HMG proteins. Since both of deiminated HMG protein still showed binding ability to dna, padase sensitive arginine residue of the hmg porteins may play a functional role in the change of superhelical structure of DNA . These results suggest that PADase may participates in the regulation of gene expression in vivo.
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