TY - JOUR
T1 - A non-denaturing gel electrophoresis system for the purification of membrane bound proteins
AU - Cavinato, Anna G.
AU - Macleod, Robert M.
AU - Ahmed, Mahmoud S.
N1 - Funding Information:
This work has been supported by National Science Foundation Grant #83-17217 to M S A. A G C was a postdoctoral fellow supported by the same grant.
PY - 1988/6/1
Y1 - 1988/6/1
N2 - A new method is described for the purification of a membrane bound glycoprotein, the kappa opioid receptor from human placental tissue. The method uses preparative slab-gel electrophoresis in the presence of the non-denaturing detergent CHAPS. A linear relationship between log molecular weight and SDS PAGE electrophoretic mobility of known molecular weight markers, in the presence of CHAPS, is observed. Using this method, we were able partially to purify an 3H-etorphine binding glycoprotein, from placental villus tissue, with an apparent molecular weight range of 60-70,000. The iodinated glycoprotein migrates in SDS PAGE with an apparent molecular weight of 63,000. This method may be useful for the isolation of membrane bound proteins, especially when an affinity ligand is not available.
AB - A new method is described for the purification of a membrane bound glycoprotein, the kappa opioid receptor from human placental tissue. The method uses preparative slab-gel electrophoresis in the presence of the non-denaturing detergent CHAPS. A linear relationship between log molecular weight and SDS PAGE electrophoretic mobility of known molecular weight markers, in the presence of CHAPS, is observed. Using this method, we were able partially to purify an 3H-etorphine binding glycoprotein, from placental villus tissue, with an apparent molecular weight range of 60-70,000. The iodinated glycoprotein migrates in SDS PAGE with an apparent molecular weight of 63,000. This method may be useful for the isolation of membrane bound proteins, especially when an affinity ligand is not available.
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U2 - 10.1080/00327488808062522
DO - 10.1080/00327488808062522
M3 - Article
C2 - 2836848
AN - SCOPUS:0023781974
SN - 0032-7484
VL - 18
SP - 205
EP - 216
JO - Preparative Biochemistry
JF - Preparative Biochemistry
IS - 2
ER -