title: G protein function in the ischaemic myocardium.
abstract: The activity of adenylyl cyclase (AC) is controlled by its interaction with receptor-regulated G proteins. The efficiency to form cyclic AMP is strongly influenced by the amount, the subspecies and function of these regulatory proteins. An impairment of AC function has been shown to occur in sarcolemmal preparations (SL) of hearts exposed to either local or global ischaemia. To examine the contribution of G protein function to this phenomenon, cholera toxin (CT)-catalysed ADP-ribosylation of Gs and pertussis toxin (PT)-catalysed ADP-ribosylation of G proteins have been investigated in SL of porcine hearts exposed to global ischaemia for 15-45 min. ADP-ribosylation by CT of an approximately 45 kDa polypeptide was 0.46 +/- 0.06 and ADP-ribosylation by PT of three 39-41 kDa polypeptides was 4.77 +/- 0.77 pmol mg-1 protein in SL of non-ischaemic myocardium. Whereas no change was observed in CT-catalyzed ribosylation after 30 min of ischaemia, there was a reduction in PT-catalyzed ADP-ribosylation to 3.7 +/- 0.35 pmol mg-1 protein after 30 min of ischaemia. Prolongation of ischaemia to 45 min did not reduce further ADP-ribosylation capacity. Quantitative immunoblotting of PT-sensitive G proteins suggests that the diminution of ADP-ribosylation occurred because of a loss of alpha-subunits of G0, Gi-1, and Gi-2 from sarcolemmal membranes.
authors.affiliation: Institute of Cardiovascular Research, Division of Cellular and Molecular Cardiology, Berlin-Buch, Germany.
authors.forename: L
authors.initials: L
authors.lastname: Will-Shahab
authors.forename: W
authors.initials: W
authors.lastname: Rosenthal
authors.forename: W
authors.initials: W
authors.lastname: Schulze
authors.forename: I
authors.initials: I
authors.lastname: Küttner
mesh_terms: D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002772:Cholera Toxin; D003327:Coronary Disease / Q000201:enzymology / Q000503:physiopathology*; D019204:GTP-Binding Proteins / Q000032:analysis*; D015151:Immunoblotting; D066298:In Vitro Techniques; D009206:Myocardium / Q000737:chemistry* / Q000201:enzymology; D037342:Pertussis Toxin; D012508:Sarcolemma / Q000737:chemistry* / Q000201:enzymology; D013552:Swine; D010566:Virulence Factors, Bordetella
chemical_list: D037361:Adenylate Cyclase Toxin; D010566:Virulence Factors, Bordetella; D000246:Adenosine Diphosphate Ribose; D002772:Cholera Toxin; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases