High-performance liquid chromatography of the main polypeptide (MP26) of lens fiber plasma membranes solubilized with n-octyl β-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 S 20,w value obtained from analytical ultracentrifugation and Stokes radius determined by HPLC of the solu...

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Bibliographic Details
Published in:FEBS letters 1988-06, Vol.233 (1), p.148-152
Main Authors: Manenti, S., Dunia, I., le Maire, M., Benedetti, E.L.
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Aquaporins
Biological and medical sciences
Cell Membrane - analysis
Chemical Phenomena
Chemistry, Physical
Chromatography, High Pressure Liquid
Detergents
Electrophoresis, Polyacrylamide Gel
eye lens
Eye Proteins - analysis
Fatty Acids - analysis
Fundamental and applied biological sciences. Psychology
Gel chromatography
Glucosides
Glycosides
Immunoassay
Lens, Crystalline - analysis
Macromolecular Substances
Membrane Glycoproteins
Membrane protein
membrane proteins
Miscellaneous
Monomer
Phosphoproteins
Proteins
Serine Endopeptidases
Solubility
Solubilization
Ultracentrifugation
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Summary: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 S 20,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 NH 2 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).
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(88)81373-5