High-performance liquid chromatography of the main polypeptide (MP26) of lens fiber plasma membranes solubilized with n-octyl beta-D-glucopyranoside.
The main polypeptide isolated from lens fiber membrane has been solubilized in octyl glucoside and studied by gel filtration in high-performance liquid chromatography (HPLC). The combination of S20,w value obtained from analytical ultracentrifugation and Stokes radius determined by HPLC of the soluble fraction indicates that more than 90% of the protein is monomeric. The solubilization of the protein seems to be dependent upon the presence of the NH2 and COOH terminal sequences, since proteolytic degradation of MP26 which removes these terminal sequences is less soluble than the uncleaved polypeptide. Moreover, there is a higher amount of oligomer after proteolysis. Fatty acid analysis by gas chromatography shows that the insoluble membrane fraction from both cortical and nuclear fibers comprises a special class of long (C22) saturated fatty acids (behenic acid).
Manenti
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Institut Jacques Monod, Centre National de la Recherche Scientifique (CNRS), Université Paris VII, France.
Dunia
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le Maire
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Benedetti
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D020346:Aquaporins; D002462:Cell Membrane / Q000032:analysis; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D005136:Eye Proteins / Q000032:analysis*; D005227:Fatty Acids / Q000032:analysis; D005960:Glucosides; D006027:Glycosides; D007118:Immunoassay; D007908:Lens, Crystalline / Q000032:analysis*; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D010750:Phosphoproteins; D012697:Serine Endopeptidases; D012995:Solubility; D014461:Ultracentrifugation
D020346:Aquaporins; D003902:Detergents; D005136:Eye Proteins; D005227:Fatty Acids; D005960:Glucosides; D006027:Glycosides; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D010750:Phosphoproteins; C030328:aquaporin 0; C018619:octyl-beta-D-glucoside; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; C007547:behenic acid