| Project/Area Number |
15590077
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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 | Kyoto Sangyo University |
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Principal Investigator |
KUROSAKA Akira Kyoto Sangyo University, Faculty of Engineering, Professor, 工学部, 教授 (90186536)
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| Co-Investigator(Kenkyū-buntansha) |
TERACHI Toru Kyoto Sangyo University, Faculty of Engineering, Professor, 工学部, 教授 (90202192)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2004: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2003: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | mucin carbohydrates / neurodegenerative disease / glycosyltransferase / α-synuclein / zebrafish / neural development / 構造活性相関 / レクチン / GalNAc転移酵素 |
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| Research Abstract |
1.Addition of mucin-type carbohydrates to a protein involved in neurodegenerative diseases
In vitro addition of GalNAc to α-synuclein, which is reported to aggregate to form insoluble fibrils in Parkinson's disease, was investigated using four GalNAc-T isozymes as enzyme source, and α-synuclein expressed in E.coli as substrate. Of four isozymes, only two isozymes transferred GalNAc to α-synuclein. Endoplasmic reticulum stress was given to cultured neural cells to see if it may cause transcriptional induction of neuron-specific GalNAc-transferases. No induction was, however, observed.
2.Function of mucin-carbohydrates during development and differentiation of nervous tissues
When expression of neuron-specific GalNAc-transferase isozymes was depressed using an RNAi method in zebrafish embryos, disorder of hindbrain development was observed. Similarly, expression of these isozymes was suppressed in pluripotent embryonic carcinoma P19 cells. Neuronal differentiation of these cells was inhibited as well.
3.Structure-function relationship of a GalNAc-transferase
Function of the lectin-like of GalNAc-T1 was investigated. GalNAc-T1 bound to resins coated with either GalNAc or Man, suggesting the sugar binding activity of the lectin domain. Moreover, study with site-directed mutagenesis indicated that one of the tryptophan residues in the Gal/GalNAc-T motif is involved in the recognition of both donor and acceptor substrates.
4.Substrate screening of neuron-specific isozymes
Phage-display screening was carried out to find peptides that can bind to the enzyme with high affinity, but no peptides have been obtained so far.
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