Lens plasma membrane: isolation and biochemical characterization.

Lens plasma membrane from different animal lenses has been prepared by the acylation procedure. Using three different criteria: heat aggregation of intrinsic membrane polypeptides, immunochemistry and solubilization of intrinsic proteins at low (40 mM) LIS concentration, it has been shown that these preparations are essentially free of cytoplasmic contaminants. Using the results obtained with acylated membrane as the reference of purity, it has been shown that both sucrose gradient centrifugation of bovine WI protein and urea washing of old human lenses give impure membrane preparations. The main intrinsic polypeptides (mol. wt 26 000 and 22 000) of human lens membrane have been purified and characterized. It has been shown by enzymatic digestion and amino terminal analysis of the residual membrane-bound fragments that the amino terminal halves of the polypeptides are embedded in the lipid bilayer and are probably blocked at their amino terminal sites. Lipid analyses of human and bovine lens membranes suggest that the protein to total lipid ratio is 1:1. Carbohydrate analyses of chromatographically separated intrinsic membrane polypeptides indicate the presence of 1 mol glucose/1 mol protein.