Purification and characterization of spiralin, the main protein of the Spiroplasma citri membrane.
The membrane proteins from Spiroplasma citri have been resolved into 16 components by SDS-polyacrylamide gel electrophoresis. By this technique it was also shown that the molecular weights of these proteins ranged from 13000 to 160 000. One of the proteins, which had an apparent molecular weight of 26 000 was the most abundant and represented more than 22% of total membrane protein. We have designated this protein spiralin. None of the proteins contained carbohydrate. Spiralin has been isolated by a procedure which involves removal of some membrane proteins with the neutral detergent Tween 20, selective solubilization of the Tween residue in DOC and fractionation of the DOC-soluble material by agarose-suspension electrophoresis. The homogeneity of spiralin was demonstrated by analytical polyacrylamide gel electrophoresis under different conditions and by crossed immunoelectrophoresis. Spiralin appeared to bind less DOC than the other membrane proteins of S. citri. This observation does not imply, however, that the binding of DOC to spiralin is weak. Spiralin was neither soluble in detergent-free buffers nor in Tween 20, which indicated that it is an intrinsic membrane protein. The amino-acid composition of spiralin was quite different from that of the membrane. Spiralin lacked methionine, histidine and tryptophan, and had a low content of glycine, leucine, tyrosine and phenylalanine, but a high content of threonine, alanine and valine.
Wróblewski
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Laboratoire de Biologie Cellulaire, Faculté des Sciences Biologiques, Complexe de Beaulieu, B.P. 25 A, 35031 Rennes, France.
Johansson
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Hjérten
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D000409:Alanine / Q000737:chemistry; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D001671:Biochemistry / Q000379:methods; D002021:Buffers; D002462:Cell Membrane / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D004586:Electrophoresis; D004587:Electrophoresis, Agar Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005998:Glycine / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D007122:Immunoelectrophoresis; D007425:Intracellular Membranes / Q000378:metabolism; D007930:Leucine / Q000737:chemistry; D008715:Methionine / Q000737:chemistry; D008970:Molecular Weight; D010649:Phenylalanine / Q000737:chemistry; D012685:Sepharose / Q000378:metabolism / Q000494:pharmacology; D045642:Spiroplasma citri / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D013912:Threonine / Q000737:chemistry; D014364:Tryptophan / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry; D014633:Valine / Q000737:chemistry
D001425:Bacterial Outer Membrane Proteins; D002021:Buffers; D003902:Detergents; C028588:spiralin; D013912:Threonine; D014443:Tyrosine; D010649:Phenylalanine; D006639:Histidine; D014364:Tryptophan; D012685:Sepharose; D008715:Methionine; D007930:Leucine; D014633:Valine; D000409:Alanine; D005998:Glycine