Use of zonal centrifugation method for the preparation of mu-opioid receptor enriched membranes from bovine corpus striatum.
Earlier attempts to purify the opioid receptors met with limited success because of the use of brain crude membranes. Subcellular fractionation procedures adopted now to get enriched membranes resulted in 2-3 fold enrichment. However, a procedure has been developed in our laboratory in which a crude membrane fraction obtained at 17,500 x g was lysed with 1 mM sodium bicarbonate and later subjected to zonal fractionation, employing a density gradient of 0.6-1.2 M sucrose. This could yield a membrane fraction highly enriched with mu-opioid receptors which is 9.3 fold higher than the crude membranes.
Sastry
K V
KV
Department of Neurochemistry, National Institute of Nutrition, Jamai Osmania, Hyderabad, India. icmrnin@ren.nic.in
Harikumar
K G
KG
Sailaja
P
P
Janardana Sarma
M K
MK
D000818:Animals; D002417:Cattle; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure*; D002501:Centrifugation, Zonal / Q000379:methods; D003342:Corpus Striatum / Q000378:metabolism / Q000648:ultrastructure*; D004174:Diprenorphine / Q000378:metabolism; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D004745:Enkephalins / Q000378:metabolism; D011869:Radioligand Assay; D017450:Receptors, Opioid, mu / Q000819:agonists / Q000302:isolation & purification* / Q000378:metabolism; D013347:Subcellular Fractions / Q000648:ultrastructure; D013395:Sucrose; D013574:Synaptosomes / Q000378:metabolism / Q000648:ultrastructure
D004745:Enkephalins; D017450:Receptors, Opioid, mu; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D004174:Diprenorphine; D013395:Sucrose