Isolation of Silk Proteins from a Caddisfly Larva, Stenopsyche Marmorata
DOI:
10.3993/jfbi06201202
Journal of Fiber Bioengineering & Informatics, 5 (2012), pp. 125-137.
Published online: 2012-05
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@Article{JFBI-5-125,
author = {},
title = {Isolation of Silk Proteins from a Caddisfly Larva, Stenopsyche Marmorata},
journal = {Journal of Fiber Bioengineering and Informatics},
year = {2012},
volume = {5},
number = {2},
pages = {125--137},
abstract = {The protein composition of the silk gland of caddisfly larva, Stenopsyche marmorata was investigated.
Dissected silk glands of the larvae were treated in a Tris buffer (pH 8.0) containing 8 M urea, and the
proteins in the lumen of the silk gland were solubilized by freeze and thaw. The major proteins of the silk
gland were precipitated upon dialysis against diluted acetic acid. The precipitate was dissolved in a basic
Tris buffer containing high concentrations of urea and dithiothreitol. Trypsin-like proteinase activity was
detected in the silk gland extract. Therefore, all extraction procedures were re-visited in the presence
of Leupeptin, which is a specific inhibitor of the trypsin-like proteinase in the silk gland. The improved
preparation of the silk gland extract contains a high molecular weight protein, namely S. marmorata
silk protein-1 (Smsp-1), of which the molecular weight was estimated as 310-320 kDa on the basis
of electrophoretic mobility on cetyltrimethylammonium bromide-poly (acryl amide) gel electrophoresis.
Subsequent gel filtration exhibited a chromatogram, where the Smsp-1 bands were found from void
(ca. 2000 kDa) toward the 300 kDa region, latter of which corresponds to the estimated molecular
weight of monomeric Smsp-1. The Smsp-1 eluted near void region was found to be electrophoretically
homogenous, suggesting that Smsp-1 molecules associate in a multimeric form. Sodium dodecylsulfate-
PAGE indicated that the fractions eluted at the 650-280 kDa region also contain Smsp-1, together with
three kinds of low molecular weight proteins, which were designated as Smsp-2 (26 kDa), -3 (21 kDa),
and -4 (16 kDa). These results suggest that the Smsps-1, -2, -3, and -4 can form a larger complex to be
spun into net threads.},
issn = {2617-8699},
doi = {https://doi.org/10.3993/jfbi06201202},
url = {http://global-sci.org/intro/article_detail/jfbi/4868.html}
}
TY - JOUR
T1 - Isolation of Silk Proteins from a Caddisfly Larva, Stenopsyche Marmorata
JO - Journal of Fiber Bioengineering and Informatics
VL - 2
SP - 125
EP - 137
PY - 2012
DA - 2012/05
SN - 5
DO - http://doi.org/10.3993/jfbi06201202
UR - https://global-sci.org/intro/article_detail/jfbi/4868.html
KW - Stenopsyche marmorata
KW - Silk gland proteins
KW - Protein composition analysis
KW - Isolation
AB - The protein composition of the silk gland of caddisfly larva, Stenopsyche marmorata was investigated.
Dissected silk glands of the larvae were treated in a Tris buffer (pH 8.0) containing 8 M urea, and the
proteins in the lumen of the silk gland were solubilized by freeze and thaw. The major proteins of the silk
gland were precipitated upon dialysis against diluted acetic acid. The precipitate was dissolved in a basic
Tris buffer containing high concentrations of urea and dithiothreitol. Trypsin-like proteinase activity was
detected in the silk gland extract. Therefore, all extraction procedures were re-visited in the presence
of Leupeptin, which is a specific inhibitor of the trypsin-like proteinase in the silk gland. The improved
preparation of the silk gland extract contains a high molecular weight protein, namely S. marmorata
silk protein-1 (Smsp-1), of which the molecular weight was estimated as 310-320 kDa on the basis
of electrophoretic mobility on cetyltrimethylammonium bromide-poly (acryl amide) gel electrophoresis.
Subsequent gel filtration exhibited a chromatogram, where the Smsp-1 bands were found from void
(ca. 2000 kDa) toward the 300 kDa region, latter of which corresponds to the estimated molecular
weight of monomeric Smsp-1. The Smsp-1 eluted near void region was found to be electrophoretically
homogenous, suggesting that Smsp-1 molecules associate in a multimeric form. Sodium dodecylsulfate-
PAGE indicated that the fractions eluted at the 650-280 kDa region also contain Smsp-1, together with
three kinds of low molecular weight proteins, which were designated as Smsp-2 (26 kDa), -3 (21 kDa),
and -4 (16 kDa). These results suggest that the Smsps-1, -2, -3, and -4 can form a larger complex to be
spun into net threads.
Kousaku Ohkawa, Yumi Miura, Takaomi Nomura, Ryoichi Arai, Koji Abe, Masuhiro Tsukada & Kimio Hirabayashi. (2019). Isolation of Silk Proteins from a Caddisfly Larva, Stenopsyche Marmorata.
Journal of Fiber Bioengineering and Informatics. 5 (2).
125-137.
doi:10.3993/jfbi06201202
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