Document Detail

Characterization of a membrane glycoprotein having pharmacological and biochemical properties of an AT2 angiotensin II receptor from human myometrium.
MedLine Citation:
PMID:  8143746     Owner:  NLM     Status:  MEDLINE    
The angiotensin II receptors of human myometrial tissue were characterized using ligand binding, cross-linking with radioactive label, detergent solubilization and partial purification by lectin-affinity chromatography. Human myometrial membrane preparations contained variable amount (5-650 fmol/mg protein) of high affinity (Kd = 44-65 pM) binding sites for 125I-CGP42112, a ligand specific for the AT2 subtype of angiotensin II receptors. Competition studies with AT1-specific and AT2-specific compounds indicated that angiotensin II receptors on these membranes were exclusively of the AT2 subtype. The binding sites for 125I-CGP42112 were efficiently solubilized by the detergent Chaps, albeit with a marked decrease in affinity (Kd = 1.2 nM). The proteins in the myometrial membrane preparation were cross-linked to 125I-[Sar1, Ile8]angiotensin II (Sarile) with disuccinimidyl suberate. When low concentrations of cross-linker were used, a single radiolabelled band of about 66-70 kDa was revealed on SDS/PAGE. At higher concentrations additional bands of about 105-120 kDa and 200 kDa were labelled. The 66-70-kDa and 105-120-kDa bands were separated by electroelution of SDS/PAGE gel slices and submitted to trypsin cleavage. The tryptic-peptide maps were identical for both products, suggesting that the additional bands are homodimers and trimers of the labelled polypeptide. The Chaps-solubilized receptor was retained on wheat-germ-agglutinin-Sepharose and specifically eluted by the competing sugar triacetylchitotriose, leading to a fivefold purification factor. Treatment of the 125I-Sarile-labelled protein with N-glycanase caused a shift in its apparent molecular mass on SDS/PAGE from 66-70 kDa to 33 kDa.
D Lazard; P Villageois; M M Briend-Sutren; F Cavaillé; S Bottari; A D Strosberg; C Nahmias
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  European journal of biochemistry / FEBS     Volume:  220     ISSN:  0014-2956     ISO Abbreviation:  Eur. J. Biochem.     Publication Date:  1994 Mar 
Date Detail:
Created Date:  1994-04-29     Completed Date:  1994-04-29     Revised Date:  2007-07-23    
Medline Journal Info:
Nlm Unique ID:  0107600     Medline TA:  Eur J Biochem     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  919-26     Citation Subset:  IM    
ICGM, Laboratoire d'Immunopharmacologie Moléculaire, CNRS UPR 0415, Paris, France.
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MeSH Terms
1-Sarcosine-8-Isoleucine Angiotensin II / metabolism
Cell Membrane / metabolism
Cross-Linking Reagents
Membrane Glycoproteins / chemistry,  metabolism
Molecular Weight
Myometrium / chemistry*
Oligopeptides / metabolism
Receptors, Angiotensin / chemistry*,  metabolism
Reg. No./Substance:
0/Cross-Linking Reagents; 0/Membrane Glycoproteins; 0/Oligopeptides; 0/Receptors, Angiotensin; 127060-75-7/CGP 42112A; 9088-01-1/1-Sarcosine-8-Isoleucine Angiotensin II

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