PMID- 6284211
OWN - NLM
STAT- MEDLINE
DCOM- 19820910
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 21
IP  - 9
DP  - 1982 Apr 27
TI  - Identification of the glucagon receptor by covalent labeling with a radiolabeled 
      photoreactive glucagon analogue.
PG  - 1996-2004
AB  - The photoreactive 125I-labeled glucagon-NAPS [125I-labeled 
      2-[2-nitro-4-azidophenyl)sulfenyl]-Trp25-glucagon] was used to label the glucagon 
      receptor sites in rat liver plasma membranes. The proteins labeled were analyzed 
      by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with or without 
      reduction with dithiothreitol. The photoaffinity peptide specifically labeled a 
      number of bands with apparent molecular weights greater than 200000 and probably 
      at least two protein bands in the molecular weight range 52000-70000. The 
      relative amounts of radioactivity associated with these bands and their relative 
      mobilities differed in samples from reduced and unreduced membranes. Their 
      relative mobilities also differed with percent acrylamide cross-linking, 
      suggesting a glycoprotein nature and the presence of intramolecular disulfide 
      bonds. A nonspecifically labeled band with an apparent molecular weight of 
      27000-28000 also displayed a similar behavior. Photolabeling in the presence of 
      0.1 mM guanosine 5'-triphosphate (GTP) decreased the amount of radiolabeling of 
      these bands, suggesting their involvement in the glucagon stimulation of 
      adenylate cyclase. The photolabeled receptor in the membranes, solubilized with 
      Lubrol-PX and fractionated on an Ultrogel AcA22 column, eluted with an apparent 
      molecular weight of 200000-250000. Addition of GTP to the solubilized glucagon 
      receptor of nonirradiated membranes caused complete dissociation of the complex. 
      Gel electrophoresis of the partially purified radiolabeled receptor identified 
      the same protein components observed in photolabeled membranes. These results 
      indicate that the glucagon receptor is an oligomer probably composed of at least 
      two different subunits that are linked together or greatly stabilized by 
      disulfide bonds. They also show that 125I-labeled glucagon-NAPS can be used 
      effectively to covalently label the putative glucagon receptor and thus aid in 
      its further characterization.
FAU - Demoliou-Mason, C
AU  - Demoliou-Mason C
FAU - Epand, R M
AU  - Epand RM
LA  - eng
GR  - AM 21285/AM/NIADDK NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Affinity Labels)
RN  - 0 (Azides)
RN  - 0 (Receptors, Cell Surface)
RN  - 0 (Receptors, Glucagon)
RN  - 76359-36-9 (glucagon, 2-nitro-4-azidophenylsulfenyl-)
RN  - 9007-92-5 (Glucagon)
SB  - IM
MH  - Affinity Labels/metabolism
MH  - Animals
MH  - Azides/*metabolism
MH  - Cell Membrane/metabolism
MH  - Glucagon/*metabolism
MH  - In Vitro Techniques
MH  - Liver/metabolism
MH  - Molecular Weight
MH  - Photochemistry
MH  - Rats
MH  - Rats, Inbred Strains
MH  - Receptors, Cell Surface/isolation & purification/*metabolism
MH  - Receptors, Glucagon
EDAT- 1982/04/27 00:00
MHDA- 1982/04/27 00:01
CRDT- 1982/04/27 00:00
PHST- 1982/04/27 00:00 [pubmed]
PHST- 1982/04/27 00:01 [medline]
PHST- 1982/04/27 00:00 [entrez]
AID - 10.1021/bi00538a004 [doi]
PST - ppublish
SO  - Biochemistry. 1982 Apr 27;21(9):1996-2004. doi: 10.1021/bi00538a004.