Molecular Heterogeneity of TIME-EA4, a Timer Protein in Silkworm Diapause Eggs
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概要
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TIME-EA4 is an ATPase that measures time intervals as a diapause-duration clock found in diapause eggs of the silkworm, Bombyx mori. In the current studies, we report the molecular heterogeneity of TIME-EA4 protein regarding not only amino acid L62V, but also the numbers and linkage patterns of the sugar chain attached to the Asn22 residue. These sugar chain structures were determined in a pico-molar amount of the protein by combining the methods of chemical modification (Smith degradation) and nano-HPLC-electrospray ionization-quadrupole-time of fight-mass spectrometry (ESI-Q-TOF-MS) and -MS/MS. The Japanese and bi-voltine Thai silkworm strains were compared to show the heterogeneity represented by four kinds of molecular species. Judicious choice of the combination methods led us to find the first example of a linkage-position difference in the glycosidic bonds even in sugar moieties of the same molecular weight; thus, the Man(1-6)Man(1-4)GlcNAc(1-4)GlcNAc structure in the C108 pure strain and Man(1-3)Man(1-4)GlcNAc(1-4)GlcNAc structure in the Kinshu-Showa hybrid. A total of five kinds of molecular heterogeneity was determined, including the amino acids in TIME-EA4 protein. This paper describes the details for determining the sugar chain linkage in TIME-EA4 from the diapause eggs of various silkworm strains.
- 2009-07-23
著者
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磯部 稔
名古屋大学高等研究院
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TANI Naoki
Laboratory of Organic Chemistry, Graduate School of Bioagricultural Sciences, Nagoya University
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磯部 稔
名古屋大学農学部
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ISOBE Minoru
Laboratory of Organic Chemistry, School of Bioagricultural Sciences, Nagoya University
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磯部 稔
名古屋大学大学院
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Tani Naoki
Laboratory Of Organic Chemistry Faculty Of Engineering Kanagawa University
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Isobe Minoru
Laboratory Of Organic Chemistry Faculty Of Agriculture Nagoya University
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KAI Hidenori
Laboratory of Insect Biochemistry, Faculty of Agriculture, Tottori University
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Kai Hidenori
Lab. Of Insect Biochemistry And Biotechnology Dep. Of Biochemistry And Biotechnology Fac. Of Agricul
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Kai Hidenori
Laboratory Of Applied Entomology Faculty Of Agriculture Tottori University
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Pitchayawasin Thapphasaraphong
Laboratory Of Organic Chemistry Graduate School Of Bioagricultural Sciences Nagoya University
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Kato Junko
Laboratory Of Organic Chemistry Graduate School Of Bioagricultural Sciences Nagoya University
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KURAHASHI Takuya
Laboratory of Organic Chemistry, Graduate School of Bioagricultural Sciences, Nagoya University
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TRISUNAN Saikwan
Chul Thai Silk Co., Ltd.
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Trisunan Saikwan
Chul Thai Silk Co. Ltd.
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Kurahashi Takuya
Laboratory Of Organic Chemistry Graduate School Of Bioagricultural Sciences Nagoya University
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Isobe Minoru
Laboratory Of Organic Chemistry Bioagricultural Sciences Nagoya University
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Pitchayawasin-Thapphasaraphong Suthasinee
Laboratory of Organic Chemistry, Graduate School of Bioagricultural Sciences, Nagoya University
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