Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
10
pubmed:dateCreated
1994-6-16
pubmed:abstractText
Active opioid binding proteins were solubilized from rat brain membranes in high yield with sodium deoxycholate in the presence of NaCl. Purification of opioid binding proteins was accomplished by opioid antagonist affinity chromatography. Chromatography using the delta-opioid antagonist N,N-diallyl-Tyr-D-Leu-Gly-Tyr-Leu attached to omega-aminododecyl-agarose (Affi-G) (procedure A) yielded a partially purified protein that binds selectively the delta-opioid agonist [3H]Tyr-D-Ser-Gly-Phe-Leu-Thr ([3H]DSLET), with a Kd of 19 +/- 3 nM and a Bmax of 5.1 +/- 0.4 nmol/mg of protein. Subsequently, Lens culinaris agglutinin-Sepharose 4B chromatography of the Affi-G eluate resulted in isolation of an electrophoretically homogeneous protein of 58 kDa that binds selectively [3H]DSLET with a Kd of 21 +/- 3 nM and a Bmax of 16.5 +/- 1.0 nmol/mg of protein. Chromatography using the nonselective antagonist 6-aminonaloxone coupled to 6-aminohexanoic acid-Sepharose 4B (Affi-NAL) (procedure B) resulted in isolation of a protein that binds selectively [3H]DSLET with a Kd of 32 +/- 2 nM and a Bmax of 12.4 +/- 0.5 nmol/mg of protein, and NaDodSO4/PAGE revealed a major band of apparent molecular mass 58 kDa. Polyclonal antibodies (Anti-R IgG) raised against the Affi-NAL protein inhibit the specific [3H]DSLET binding to the Affi-NAL eluate and to the solubilized membranes. Moreover, the Anti-R IgG inhibits the specific binding of radiolabeled Tyr-D-Ala-Gly-N-methyl-Phe-Gly-ol (DAMGO; mu-agonist), DSLET (delta-agonist), and naloxone to homogenates of rat brain membranes with equal potency. Furthermore, immunoaffinity chromatography of solubilized membranes resulted in the retention of a major protein of apparent molecular mass 58 kDa. In addition, immunoblotting of solubilized membranes and purified proteins from the Affi-G and Affi-NAL matrices revealed that the Anti-R IgG interacts with a protein of 58 kDa.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-1334555, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-1335167, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-1335327, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-2156011, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-2160790, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-2552439, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-2826227, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-2884604, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-2991902, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-2993292, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-2999106, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-3012554, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-3037220, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-3843705, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-4117461, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-5432063, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-6095896, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-6175245, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-6254391, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-6301606, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-6312839, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-6313043, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-6319079, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-7509520, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-8392341, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-8396046, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-894682, http://linkedlifedata.com/resource/pubmed/commentcorrection/8183950-942051
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0027-8424
pubmed:author
pubmed:issnType
Print
pubmed:day
10
pubmed:volume
91
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
4574-8
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed-meshheading:8183950-Analgesics, pubmed-meshheading:8183950-Animals, pubmed-meshheading:8183950-Antibodies, pubmed-meshheading:8183950-Binding, Competitive, pubmed-meshheading:8183950-Blotting, Western, pubmed-meshheading:8183950-Brain, pubmed-meshheading:8183950-Cell Membrane, pubmed-meshheading:8183950-Chromatography, Affinity, pubmed-meshheading:8183950-Chromatography, Gel, pubmed-meshheading:8183950-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:8183950-Enkephalin, Ala(2)-MePhe(4)-Gly(5)-, pubmed-meshheading:8183950-Enkephalin, Leucine, pubmed-meshheading:8183950-Enkephalins, pubmed-meshheading:8183950-Kinetics, pubmed-meshheading:8183950-Male, pubmed-meshheading:8183950-Molecular Weight, pubmed-meshheading:8183950-Oligopeptides, pubmed-meshheading:8183950-Rats, pubmed-meshheading:8183950-Rats, Wistar, pubmed-meshheading:8183950-Receptors, Opioid, delta, pubmed-meshheading:8183950-Ultrafiltration
pubmed:year
1994
pubmed:articleTitle
Purification to homogeneity of an active opioid receptor from rat brain by affinity chromatography.
pubmed:affiliation
Institute of Biology, National Center for Scientific Research Demokritos, Aghia Paraskevi Attiki, Greece.
pubmed:publicationType
Journal Article