Purification and characterization of fibroin from the tropical Saturniid silkworm, Antheraea mylitta

被引：45

作者：

Datta, A ^{[1
]}

Ghosh, AK ^{[1
]}

Kundu, SC ^{[1
]}

机构：

[1] Indian Inst Technol, Dept Biotechnol, Kharagpur 721302, W Bengal, India

来源：

INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY | 2001年 / 31卷 / 10期

关键词：

Antheraea mylitta; fibroin; two-dimensional electrophoresis; homodimer; deglycosylation; O-glycosylation;

D O I：

10.1016/S0965-1748(01)00049-2

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The fibroin protein isolated from the posterior silkgland of the tropical Saturniid silkworm Antheraea mylitta, was solubilized in lithium dodecyl sulfate and purified by get filtration. The major fraction from gel filtration was analyzed by SDS-PAGE under non-reducing and reducing conditions. One major protein band of ca 395 kDa was obtained under non-reducing conditions and a doublet band of similar to 197 kDa under reducing conditions. The appearance of a single spot in two-dimensional electrophoresis confirmed the purity of the protein indicating that it may be a homodimeric protein of two similar sized polypeptides. Amino acid composition analysis showed that, like other Saturniid fibroins, it is rich in glycine, alanine and serine amino acids. N-terminal amino acid sequence shows significant homology with other Antheraea species. The enzymatic deglycosylation analysis indicates that the fibroin protein is glycosylated and the oligosaccharides are O-linked to the protein back-bone by N-acetylgalactoseamine moiety which conforms to a Core 1 mucin-type glycosylation pattern. (C) 2001 Elsevier Science Ltd. All rights reserved.

引用

页码：1013 / 1018

页数：6

共 20 条

[11] FURTHER EVIDENCE FOR IMPORTANCE OF THE SUBUNIT COMBINATION OF SILK FIBROIN IN ITS EFFICIENT SECRETION FROM THE POSTERIOR SILK GLAND-CELLS [J].

TAKEI, F ;

KIKUCHI, Y ;

KIKUCHI, A ;

MIZUNO, S ;

SHIMURA, K .

JOURNAL OF CELL BIOLOGY, 1987, 105 (01) :175-180

[12] ISOLATION AND CHARACTERIZATION OF FIBROIN MESSENGER-RNAS FROM THE SATURNIID SILKWORMS, ANTHERAEA-YAMAMAI, ANTHERAEA-PERNYI AND PHILOSAMIA-CYNTHIA-RICINI [J].

TAMURA, T ;

SAKATE, S .

INSECT BIOCHEMISTRY, 1988, 18 (02) :169-175

[13] THE FIBROIN GENES OF ANTHERAEA-YAMAMAI AND BOMBYX-MORI ARE DIFFERENT IN THEIR CORE REGIONS BUT REVEAL A STRIKING SEQUENCE SIMILARITY IN THEIR 5' ENDS AND 5' FLANKING REGIONS [J].

TAMURA, T ;

INOUE, H ;

SUZUKI, Y .

MOLECULAR & GENERAL GENETICS, 1987, 207 (2-3) :189-195

[14] Hydrophobic interaction of P25, containing Asn-linked oligosaccharide chains, with the H-L complex of silk fibroin produced by Bombyx mori [J].

Tanaka, K ;

Inoue, S ;

Mizuno, S .

INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, 1999, 29 (03) :269-276

[15] CLUSTAL-W - IMPROVING THE SENSITIVITY OF PROGRESSIVE MULTIPLE SEQUENCE ALIGNMENT THROUGH SEQUENCE WEIGHTING, POSITION-SPECIFIC GAP PENALTIES AND WEIGHT MATRIX CHOICE [J].

THOMPSON, JD ;

HIGGINS, DG ;

GIBSON, TJ .

NUCLEIC ACIDS RESEARCH, 1994, 22 (22) :4673-4680

[16] ELECTROPHORETIC TRANSFER OF PROTEINS FROM POLYACRYLAMIDE GELS TO NITROCELLULOSE SHEETS - PROCEDURE AND SOME APPLICATIONS [J].

TOWBIN, H ;

STAEHELIN, T ;

GORDON, J .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1979, 76 (09) :4350-4354

[17]

Van den Steen P, 1998, CRIT REV BIOCHEM MOL, V33, P151

[18] STRUCTURE OF A PROTEIN SUPERFIBER - SPIDER DRAGLINE SILK [J].

XU, M ;

LEWIS, RV .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (18) :7120-7124

[19] Preferential codon usage and two types of repetitive motifs in the fibroin gene of the Chinese oak silkworm, Antheraea pernyi [J].

Yukuhiro, K ;

Kanda, T ;

Tamura, T .

INSECT MOLECULAR BIOLOGY, 1997, 6 (01) :89-95

[20] Fine organization of Bombyx mori fibroin heavy chain gene [J].

Zhou, CZ ;

Confalonieri, F ;

Medina, N ;

Zivanovic, Y ;

Esnault, C ;

Yang, T ;

Jacquet, M ;

Janin, J ;

Duguet, M ;

Perasso, R ;

Li, ZG .

NUCLEIC ACIDS RESEARCH, 2000, 28 (12) :2413-2419

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