Disassembly and reassembly in vitro of complexes of secretory proteins from Chironomus tentans salivary glands.
The secretory proteins of Chironomus tentans larvae form insoluble fibers that are spun into threads used to construct underwater feeding and pupation tubes. We began in vitro studies of the mechanism of assembly into fibers, the structure of the assembled proteins, and the contribution of individual proteins to the assembled structure. From measurements of turbidity and electron micrographs, we observed that the secretory proteins were isolated as complexes. These complexes are most likely at initial stages of assembly; further assembly into insoluble fibers must occur in vivo. Denaturation and reduction disrupted the complexes, and removal of the denaturing and reducing agents resulted in reassembly of the complexes. The circular dichroic spectrum of the complexes indicated that the assembled proteins had the tertiary structure alpha + beta. The largest secretory proteins were purified and shown to have both similar morphology, using electron microscopy, and a similar dichroic spectrum to that of the native complexes. We concluded that the large secretory proteins form the fibrous backbone of the complexes that we observe.
Wellman
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Department of Biochemistry, University of Mississippi Medical Center, Jackson 39216-4505.
Case
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D000818:Animals; D002683:Chironomidae / Q000378:metabolism*; D002942:Circular Dichroism; D004175:Diptera / Q000378:metabolism*; D008854:Microscopy, Electron; D008970:Molecular Weight; D011487:Protein Conformation; D011489:Protein Denaturation; D012469:Salivary Glands / Q000378:metabolism; D012471:Salivary Proteins and Peptides / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure
D012471:Salivary Proteins and Peptides