Hydrodynamic characterization of the major intrinsic protein from the bovine lens fiber membranes. Extraction in n-octyl-beta-D-glucopyranoside and evidence for a tetrameric structure.

Abstract

The major protein from the bovine lens fiber cell membranes, the 26-kilodalton protein (major intrinsic protein (MIP26)), has been solubilized in n-octyl-beta-D-glucopyranoside and purified by gel filtration. The final preparation was free of detergent micelles. Gel electrophoresis in denaturing conditions has confirmed the purity of the protein sample. A s20,w of 5.55 S, obtained from analytical ultracentrifugation, and a D20,w of 3.62 x 10(-7) cm2 s-1, obtained from photon correlation spectroscopy, resulted in a molar mass of (176,000 +/- 15,000) g/mol for the protein-detergent complex using the Svedberg relation. The measured detergent content of 0.71 g of detergent/g of protein resulted in a calculated partial specific volume of 0.787 cm3/g for the protein-detergent complex and a molar mass of 103,000 g/mol for the protein moiety. This allowed us to conclude that the protein-detergent complex contains four copies of the MIP26 protein, which supports the suggestion that in vivo the MIP26 molecules cluster in tetramers to form a pore-like structure.