Cited by 784Silk-based biomaterials and applicationsSilkworms and Sericulture ResearchProtein purification and stability

Silk fibroin of Bombyx mori is secreted, assembling a high molecular mass elementary unit consisting of H-chain, L-chain, and P25, with a 6:6:1 molar ratio

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Inoue, Satoshi, Tanaka, Kazunori, Arisaka, Fumio, Kimura, Sumiko, Ohtomo, Kohei, Mizuno, Shigeki·2000·Journal of Biological Chemistry

Abstract

Silk fibroin produced by the silkworm Bombyx mori consists of a heavy chain, a light chain, and a glycoprotein, P25. The heavy and light chains are linked by a disulfide bond, and P25 associates with disulfide-linked heavy and light chains by noncovalent interactions. Quantitative enzyme-linked immunosorbent assay revealed that molar ratios of the heavy chain, light chain, and P25 were 6:6:1, both in cocoons and in fibroin secreted into the lumen of posterior silk gland. Trace amounts of fibroin produced by three "naked pupa" mutants of B. mori lacked the light chain, but the molar ratio of heavy chain and P25 was also 6:1. Gel filtration chromatography and sedimentation equilibrium analysis demonstrated that a large protein complex of approximately 2.3 MDa, designated an elementary unit of fibroin having 6:6:1 molar ratios of the heavy chain, light chain, and P25, existed in posterior silk gland cells. Inaccessibility of biotinylated concanavalin A to the native elementary unit and partial dissociation of the elementary unit after incubation with excess N-glycosidase F or endoglycosidase H suggest that a single molecule of P25 is located internally and plays an important role in maintaining integrity of the complex.

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  1. Wang, J. L., Cunningham, B. A., and Edelman, G. M. (1971) Proc. Natl. Acad. Sci. U. S. A. 68, 1130 -1134
  2. Bornemann, K. D., Brewer, J. W., Beck-Engeser, G. B., Corley, R. B., Haas, I. G., and Jack, H.-M. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 4912-4916
  3. Applied Biosystems (1985) Applied Biosystems User Bulletin 14, p. 23, Applied Biosytems, Inc., Foster City, CA

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Authors
Inoue, S., Tanaka, K., Arisaka, F., Kimura, S., Ohtomo, K., & Mizuno, S.
Affiliations
  • Tohoku University (JP)
  • Tokyo Institute of Technology (JP)
  • Chiba University (JP)
Corresponding author
Mizuno, Shigeki
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2000-12
Volume
275
Issue
51
Pages
40517-40528
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2120240537
Cited by
784
References
40
Citations by year
2026: 46 · 2025: 49 · 2024: 69 · 2023: 43 · 2022: 57 · 2021: 53 · 2020: 62 · 2019: 39 · 2018: 36 · 2017: 28 · 2016: 32 · 2015: 45 · 2014: 41 · 2013: 25 · 2012: 24
FWCI
3.98
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Topics
  • Physical Sciences › Materials Science › Biomaterials › Silk-based biomaterials and applications
  • Life Sciences › Agricultural and Biological Sciences › Insect Science › Silkworms and Sericulture Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
Keywords
silk fibroin, Bombyx mori, heavy chain, light chain, molar ratio, posterior silk gland, disulfide bonds, non-covalent interactions, protein complexes, N-glycosidase F, endoglycosidase H, concanavalin A, glycosylation
MeSH terms
Amino Acid Sequence, Animals, Base Sequence, Disulfides, DNA, Electrophoresis, Polyacrylamide Gel, Fibroins, Glycoproteins, Molecular Sequence Data, Molecular Weight, Oligosaccharides, Protein Conformation, Bombyx, Insect Proteins, Silk

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