TY - JOUR
T1 - G-proteins in guinea pig airway smooth muscle
T2 - Identification and functional involvement
AU - Joshi, Sharmishtha
AU - Abebe, Worku
AU - Agrawal, Devendra K.
PY - 1996/3
Y1 - 1996/3
N2 - In airway smooth muscle, G-proteins have not been identified directly as yet. This study was an attempt to detect various types of conventional G(i)- and G(s)-proteins in purified membranes of guinea pig airway smooth muscle and to assess the involvement of the G-proteins in agonist-induced contractile response of the smooth muscle. Immunoblotting using AS/7 antibody which recognizes G(i-1/2) demonstrated the presence of polypeptides of M(r) 34, 41 and 75 kDa, Polypeptides of M(r) 43, 46 and 48 kDa were identified with RM/1 antibody that detects G(s)-type G-proteins. The AG/1 antibody that recognizes α-subunits common to all G(i)- and G(s) proteins detected two polypeptides of M(r) 41 and 75 kDa. Heterotrimeric structure of the G-proteins was confirmed by the identification of a single dense β-subunit band at M(r) 39 kDa with SW/1 antibody. Pertussis toxin (PT) ADP-ribosylated three G(i)α polypeptides of M(r) 41, 43, and 62 kDa, On the other hand, cholera toxin (CT) catalysed the ADP-ribosylation of two G(s)α polypeptides of M(r) 46 and 62 kDa. Both PT and CT attenuated the maximum contractile responses of the airway smooth muscle to the muscarinic agonist, methacholine. Pretreatment of the tissues with the sulphydryl alkylating G-protein inhibitor, N-ethylmaleimide, also inhibited the maximum contractions to methacholine. These data suggest that plasma membranes of guinea pig airway smooth muscle contain a variety of conventional, including G(s) and G(i), and other types of G-proteins, and at least a portion of the proteins present may be involved in mediating the contractile responses of the smooth muscle to an agonist such as methacholine.
AB - In airway smooth muscle, G-proteins have not been identified directly as yet. This study was an attempt to detect various types of conventional G(i)- and G(s)-proteins in purified membranes of guinea pig airway smooth muscle and to assess the involvement of the G-proteins in agonist-induced contractile response of the smooth muscle. Immunoblotting using AS/7 antibody which recognizes G(i-1/2) demonstrated the presence of polypeptides of M(r) 34, 41 and 75 kDa, Polypeptides of M(r) 43, 46 and 48 kDa were identified with RM/1 antibody that detects G(s)-type G-proteins. The AG/1 antibody that recognizes α-subunits common to all G(i)- and G(s) proteins detected two polypeptides of M(r) 41 and 75 kDa. Heterotrimeric structure of the G-proteins was confirmed by the identification of a single dense β-subunit band at M(r) 39 kDa with SW/1 antibody. Pertussis toxin (PT) ADP-ribosylated three G(i)α polypeptides of M(r) 41, 43, and 62 kDa, On the other hand, cholera toxin (CT) catalysed the ADP-ribosylation of two G(s)α polypeptides of M(r) 46 and 62 kDa. Both PT and CT attenuated the maximum contractile responses of the airway smooth muscle to the muscarinic agonist, methacholine. Pretreatment of the tissues with the sulphydryl alkylating G-protein inhibitor, N-ethylmaleimide, also inhibited the maximum contractions to methacholine. These data suggest that plasma membranes of guinea pig airway smooth muscle contain a variety of conventional, including G(s) and G(i), and other types of G-proteins, and at least a portion of the proteins present may be involved in mediating the contractile responses of the smooth muscle to an agonist such as methacholine.
KW - ADP-ribosylation
KW - Bacterial toxins
KW - G-proteins
KW - Guinea pig trachealis
KW - Immunoblotting
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U2 - 10.1006/phrs.1996.0027
DO - 10.1006/phrs.1996.0027
M3 - Article
C2 - 8880891
AN - SCOPUS:0030108647
SN - 1043-6618
VL - 33
SP - 195
EP - 202
JO - Pharmacological Research
JF - Pharmacological Research
IS - 3
ER -