In vitro folding of synthetic glycoprotein
| Project/Area Number |
18580107
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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 |
Bioproduction chemistry/Bioorganic chemistry
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| Research Institution | Tokai University |
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Principal Investigator |
HOJO Hironobu Tokai University, School of Engineering, Professor (00209214)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAHARA Yoshiaki Tokai Univeristy, School of Engineering, Professor (50087574)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,090,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥390,000)
Fiscal Year 2007: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Glvcoorotein / Glvcopeptide thioester / Segment condensation / Folding / Chemical synthesis / フォールデノング |
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| Research Abstract |
Glycoproteins are engaged in many biological processes in living organism, such as cell growth, tumor metastasis and so on. To understand the precise role of glycoproteins, it is essential to obtain chemically homogeneous glyoproteins. To this end, we have been establishing a facile method for glycoprotein synthesis based on our thioester segment condensation method. A problem remained in this method is how to fold the synthetic glycoproteins. We sometimes observed misfolding of the synthetic glycoprotein in the case of the one having short carbohydrate chain. In this research, we aimed to synthesize glycoprotein having larger carbohydrate chain, such as complex type N-linked sugar, and fold it to a correct three dimensional structure.
First, Fmoc-Asn carrying N-linked undecasaccharide unit was prepared for the solid-phase peptide synthesis (SPPS) of glycoprotein. However, we could not obtain enough amount for SPPS. Thus, we used commercially available Fmoc-Asn carring nonasaccharide unit for the synthesis of glyoprotein, which was not folded properly, when shorter sugar was attached. The result showed that even with the longer sugar chain, the glycoprotein did not fold in a particular conformation. We further synthesized nona-, mono- and nonglycosylated chemokine CCL27 composed of 95 amino acid residues using our strategy. The folding of this protein showed that the nona and monoglycosylated CCL27 had different disulfide bond pattern with other chemokines prepared previously. In contrast, nonglycosylated CCL27 had the same disulfide bonds with other chemokines. From these results, we concluded that glycosylation has significant effect on folding of polypeptide, regardless of the length of the carbohydrate chain.
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Report
(3 results)
Research Products
(42 results)
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[Journal Article] Purification and Substrate Specificity of UDP-D-xylose : 8-D-Glucoside-l, 3-D-Xylosyltransferase Involved in the Biosynthesis of the Xyl1-3Xy11-3GIcll1-O-Ser on Epidermal Growth Factor-like Domains2007
Author(s)
T., Ishimizu, K., Sano, T., Uchida, H., Teshima, K., Omichi, H., Hojo, Y., Nakahara, S., Hase
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Journal Title
J. Biochem 141
Pages: 593-600
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] Peptide Thioester Synthesis via an Auxiliary-Mediated N-S Acyl Shift Reaction in Solution2007
Author(s)
K., Nakamura, H., Mori, T., Kawakami, H., Hojo, Y., Nakahara, S., Aimoto
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Journal Title
Mt. J. Pept. Res. Ther 13
Pages: 191-202
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Synthesis of glycopeptide dendrimer by a convergent method2007
Author(s)
C., Ozawa, H., Hojo, Y., Nakahara, H., Katayama, K., Nabeshima, T., Akahane, Y., Nakahara
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Journal Title
Tetrahedron 63
Pages: 9685-9690
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Synthesis of a glycosylated peptide thioester by the Boc strategy and its application to segment condensation2006
Author(s)
E., Haginoya, H., Hojo, Y., Nakahara, Y., Nakahara, K., Nabeshima, B. P., Toole, Y, Watanabe
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Journal Title
Biosci. Biotechnol. Biochem 70
Pages: 1338-1349
NAID
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Emmprin(basigin/CD147) : matrix metalloproteinase modulator and multifunctional cell recognition molecule that plays a critical role in cancer progression2006
Author(s)
K., Nabeshima, H., Iwasaki, K., Koga, H., Hojo, J., Suzumiya, M., Kikuchi
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Journal Title
Pathol. Mt 56
Pages: 359-367
NAID
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] An Efficient Microwave-Assisted Tandem N to S Acyl Transfer and Thioester Exchange for the Preparation of Glycosylated Peptide Thioester2006
Author(s)
F., Nagaike, Y., Onuma, C., Kanazawa, H., Hojo, A., Ueki, Y., Nakahara, Y., Nakahara
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Journal Title
Org. Lett 8
Pages: 4465-4468
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Synthesis of Glycopeptide Dendrimer by the Thioester Method2007
Author(s)
H., Hojo, C., Ozawa, Y., Nakahara, H., Katayama, K., Nabeshima, T., Akahane, Y., Nakahara
Organizer
4'h International Peptide Symposium
Place of Presentation
Cairns, Australia
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Micorwave-assisted Post-SPPS thioesterification2006
Author(s)
Y., Onuma, F., Nagaike, H., Hojo, A., Ueki, Y., Nakahara, Y., Nakahara
Organizer
43'h Japanese Peptide Symposium-4th Peptide Engineering Meeting
Place of Presentation
Yokohama. Japan
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Preparation of heterodimeric glycopeptide via a ligation of glycopeptide with intein-mediated peptide thioester2006
Author(s)
H., Hojo, H., Kodaira, H., Nagasawa, T., Takahashi, Y., Nakahara
Organizer
43'h Japanese Peptide Symposium-4'h Peptide Engineering Meeting
Place of Presentation
Yokohama, Japan
Description
「研究成果報告書概要(欧文)」より
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[Presentation] N-S Acyl reaction on peptide backbone and its application to preparation of peptide thioesters2006
Author(s)
K., Nakamura, T., Kawakami, H., Hojo, Y., Nakahara, S., Aimoto
Organizer
43'h Japanese Peptide Symposium-4'h Peptide Engineering Meeting
Place of Presentation
Yokohama, Japan
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Semisynthesis of glycopeptide hormone precursor using chemical ligation of recombinant peptide thioester2006
Author(s)
H., Hojo, H., Kodaira, T., Takahashi, H., Nagasawa, Y., Nakahara
Organizer
29'h European Peptide Symposium
Place of Presentation
Gdansk, Poland
Description
「研究成果報告書概要(欧文)」より
Related Report
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