TY - JOUR
T1 - Phase separation of rat intestinal brush border membrane proteins using Triton X-114
AU - Tiruppathi, Chinnaswamy
AU - Alpers, David H.
AU - Seetharam, Bellur
N1 - Funding Information:
’ This work was supported by Grant AM 26638 from National Institutes of Health. * Abbreviations used: SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; Hepes, 4-(2-hydroxyethyl) 1-piperazineethanesulfonic acid, PMSF, phenylmethylsulfonyl fluoride; DPP, detergent-poor phase; DRP, detergent-rich phase; IF-cbl, intrinsic factor-cobalamin.
PY - 1986/3
Y1 - 1986/3
N2 - Rat intestinal microvillus membrane contains at least 24 polypeptides, of which 18 can be solubilized using Triton X-114 at 4°C. Upon phase separation at 32°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, γ-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.
AB - Rat intestinal microvillus membrane contains at least 24 polypeptides, of which 18 can be solubilized using Triton X-114 at 4°C. Upon phase separation at 32°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, γ-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.
KW - Triton X-114
KW - electrophoresis
KW - intestine
KW - membrane
KW - phase separation
KW - proteins
UR - http://www.scopus.com/inward/record.url?scp=0022478689&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(86)90100-4
DO - 10.1016/0003-2697(86)90100-4
M3 - Article
C2 - 3010762
AN - SCOPUS:0022478689
VL - 153
SP - 330
EP - 335
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
IS - 2
ER -