Phase separation of rat intestinal brush border membrane proteins using Triton X-114

✍ Scribed by Chinnaswamy Tiruppathi; David H. Alpers; Bellur Seetharam

• Publisher: Elsevier Science
• Year: 1986
• Tongue: English
• Weight: 901 KB
• Volume: 153
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(86)90100-4

✦ Synopsis

Rat intestinal microvillus membrane contains at least 24 polypeptides, of which 18 can be solubilized using Triton X-114 at 4 degrees C. Upon phase separation at 32 degrees C, 11 proteins separated nearly completely into the detergent-rich phase, while 9 proteins were found exclusively in the aqueous phase. Enzymes which were uniquely included in the detergent phase were alkaline phosphatase, leucine aminopeptidase, gamma-glutamyl transpeptidase, and Ca2+-Mg2+ ATPase. The proteins which were excluded from the detergent phase and found exclusively in the aqueous phase included the disaccharidases (glucoamylase, sucrase-isomaltase, trehalase, lactase) and the ileal receptor for the intrinsic factor-cobalamin complex. Integral membrane proteins can thus be separated during solubilization into two groups prior to further purification or characterization.