2003 Fiscal Year Final Research Report Summary
Studies on mechanism of complex formation and structure analysis of monoclonal immunoglobulins bound to lactate dehydrogenase
| Project/Area Number |
13672416
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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 |
Laboratory medicine
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| Research Institution | Shinshu University |
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Principal Investigator |
KAMEKO Fumiko SHINSHU UNIVERSITY, DEP. OF BIOMEDICAL LAB., SCHOOL OF HEALTH SCIENCE, Research Assistant, 医学部, 助手 (60126670)
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| Co-Investigator(Kenkyū-buntansha) |
SAKURABAYASHI Ikunosuke Jichi Medical School, 1st Integrated Medicine, Omiya Medical Center, Professor, 大宮医療センター・総合医学講座I, 教授 (80049056)
FUJITA Kiyotaka SHINSHU UNIVERSITY, DEP. OF BIOMEDICAL LAB., SCHOOL OF HEALTH SCIENCE, Associate Professor, 医学部, 助教授 (90313866)
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| Project Period (FY) |
2001 – 2003
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| Keywords | Lactate dehydrogenase / Bence Jones protein / NAD^+ binding site / LDH-BJP complex / Complex formation |
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| Research Abstract |
Immunochemical analysis of λ type Bence Jones protein (BJP) which showed non-specific binding with lactate dehydrogenase (LDH, EC 1.1.1.27) was carried out in a patient. The patient's BJP molecules as a dimer without 2-mercaptoethanol (2-ME), was showed a three bands, even though the monomer BJP with 2-ME was a single band by immunostaining using antiserum against λ chain after SDS-PAGE. When the purified LDH mixed with NADH was eluted through the CNBr-Sepharose 4B coupled to the patient BJP, the affinity with the adsorbent was not demonstrated. Moreover, the LD-BJP complexes were dissociated by passing the mixture of the patient BJP and the purified LDH through a column containing 5'-AMP-Sepharose 4B, which can be considered as a fragment of NAD^+. The second amino acid residue of N-terminal in the patient BJP was determined as tyrosine of the aromatic amino acid by primary structure analysis. By counter affinity electrophoresis, it was identified that LDH bound to synthetic peptide consisting of 15 amino acid residues of N-terminal which had the same structure of the patient BJP. From these facts, it seems probable that LDH combines with BJP molecule at the NAD^+ binding site producing the three dimensional structure similar to NAD^+.
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