Characterization of the protein components of Nephila clavipes dragline silk.
Spider silk is predominantly composed of structural proteins called spider fibroins or spidroins. The major ampullate silk that forms the dragline and the cobweb's frame threads of Nephila clavipes is believed to be a composite of two spidroins, designated as Masp 1 and 2. Specific antibodies indeed revealed the presence of Masp 1 and 2 specific epitopes in the spinning dope and solubilized threads. In contrast, sequencing of specific peptides obtained from solubilized threads or gland urea extracts were exclusively homologous to segments of Masp 1, suggesting that this protein is more abundantly expressed in silk than Masp 2. The strength of immunoreactivities corroborated this finding. Polypeptides reactive against both Masp 1 and 2 specific antibodies were found to be expressed in the epithelia of the tail and different gland zones and accumulated in the gland secreted material. Both extracts of gland secretion and solubilized threads showed a ladder of polypeptides in the size range of 260-320 kDa in gel electrophoresis under reducing conditions, whereas gel filtration chromatography yielded molecular masses of the proteins of approximately 300-350 kDa. In the absence of a reducing agent, dimeric forms of the spidroins were observed with estimated molecular masses of 420-480 kDa according to gel electrophoresis and 550-650 kDa as determined by gel filtration chromatography. Depending on the preparation, some silk material readily underwent degradation, and polypeptides down to 20 kDa in size and less were detectable.
Sponner
Alexander
A
Department of Biochemistry, Institute for Molecular Biotechnology, Beutenbergstrasse 11, 07745 Jena, Germany.
Schlott
Bernhard
B
Vollrath
Fritz
F
Unger
Eberhard
E
Grosse
Frank
F
Weisshart
Klaus
K
D000818:Animals; D003429:Cross Reactions; D005088:Exocrine Glands / Q000276:immunology / Q000378:metabolism; D005260:Female; D005349:Fibroins / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D007106:Immune Sera / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D019476:Insect Proteins / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D020539:Sequence Analysis, Protein; D047011:Silk / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism; D012995:Solubility; D013112:Spiders / Q000737:chemistry*
D007106:Immune Sera; D019476:Insect Proteins; D010455:Peptides; D047011:Silk; C086087:spidroin 1; C076815:spidroin 2; D005349:Fibroins