2001 Fiscal Year Final Research Report Summary
Research for the relationship of hereditary sideroblastic anemia with enzyme complex formation in mitochondria
| Project/Area Number |
12670129
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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 |
Pathological medical chemistry
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| Research Institution | Tohoku University |
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Principal Investigator |
FURUYAMA Kazumichi Tohoku University, Graduate School of Medicine, Research Associate, 大学院・医学系研究科, 助手 (80280874)
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| Co-Investigator(Kenkyū-buntansha) |
HARIGAE Hideo Tohoku University, University Hospital, Research Associate, 医学部・附属病院, 助手 (50302146)
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| Project Period (FY) |
2000 – 2001
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| Keywords | 5-aminolevulinate synthase / Succinyl CoA synthethase / Hereditary Sideroblastic Anemia |
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| Research Abstract |
1. To examine whether Erythroid specific 5-aminolevulinate synthase (ALAS-E) and ATP-specific succinyl CoA synthethase (A-SCS) form functional enzyme complex in vitro, we have expressed recombinant proteins of components of each enzyme (ALAS-E, SCS-α, SCS-βA and SCS-βG) using Baclo-virus based expression system. In this experiment, we have successfully purified enzymatically active ALAS-E recombinant protein, however, enzymatic activity of SCS could not be detected in any combination of purified component (SCS-α and SCS-βA = A-SCS, or SCS-α and SCS-βG = G-SCS). These results suggest that phosphorilation or glycosilation SCS might be needed for enzymatically active SCS, which specifically occur in only mammalian cells. Alternatively, mammalian SCS may request specific co-factor(s) for its catalytic activity. If such co-factor(s) exist, decrease amount of the co-factor may cause sideroblastic anemia.
2. To determine the specific region for interaction of ALAS-E and SCS-βA, several deletion mutants of ALAS-E protein were made. Then, the interaction of such mutant ALAS-E proteins and SCS-βA protein were determined using yeast two hybrid system. As a results, 147 amino acid deletion of N-terminal or 50 amino acid deletion of C-terminal of ALAS-E disrupt the interaction of ALAS-E and SCS-βA protein. Since these deletion mutant of ALAS-E results the disruption of homo-dimer formation of ALAS-E, homodimer formation of ALAS-E might be important for enzyme complex formation of ALAS-E and A-SCS in mitochondria.
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