Identification of guanine nucleotide binding regulatory proteins in bovine tracheal smooth muscle

Sharmishtha Joshi, Worku Abebe, Devendra K. Agrawal

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The identity of G-proteins in airway smooth muscle is not well elucidated. In the present study, by immunoblotting using AS/7 antibody specific for G(iα-1/2), EC/2 antibody specific for G(iα-3) and RM/1 antibody specific for G(sα), we identified, respectively, Mr 39, 41, 46 and 52 KDa, Mr 41 and 43 KDa, and Mr 43 and 46 KDa polypeptides of conventional (heterotrimeric) G-proteins in purified membranes of bovine tracheal smooth muscle. The identity of the Mr 41, 43 and 52 KDa G(iα), and the Mr 43 and 46 G(sα) was also confirmed by ADP-ribosylation with pertussis and cholera toxins, respectively. In addition, the common antibody (AG/1) for both G(iα) and G(sα) revealed the presence of all the above polypeptides, except the Mr 52 KDa band. By nitrocellulose blot overlay with [35S]GTPγS, we also detected seven low molecular weight GTP-binding proteins of Mr 18-30 KDa in the bovine tracheal smooth muscle. Photoaffinity crosslinking of [α-32P]GTP demonstrated the presence of high molecular GTP-binding proteins of Mr 55, 75 and 110 KDa. It is concluded that plasma membranes of bovine tracheal smooth muscle contain various types of conventional, low molecular weight and high molecular weight G-proteins. This warrantes further attention to elucidate the functional roles of G-proteins in airway smooth muscle.

Original languageEnglish (US)
Pages (from-to)179-184
Number of pages6
JournalMolecular and Cellular Biochemistry
Volume154
Issue number2
DOIs
StatePublished - Feb 9 1996

Fingerprint

Guanine Nucleotides
GTP-Binding Proteins
Smooth Muscle
Muscle
Carrier Proteins
Antibodies
Molecular Weight
Molecular weight
Proteins
Heterotrimeric GTP-Binding Proteins
Peptides
Collodion
Cholera Toxin
Pertussis Toxin
Cell membranes
Guanosine Triphosphate
Immunoblotting
Crosslinking
Adenosine Diphosphate
Cell Membrane

Keywords

  • ADP-ribosylation
  • Airway smooth muscle
  • G-proteins
  • Immunoblotting
  • Nitrocellulose blot overlay
  • Photolabeling

ASJC Scopus subject areas

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Identification of guanine nucleotide binding regulatory proteins in bovine tracheal smooth muscle. / Joshi, Sharmishtha; Abebe, Worku; Agrawal, Devendra K.

In: Molecular and Cellular Biochemistry, Vol. 154, No. 2, 09.02.1996, p. 179-184.

Research output: Contribution to journalArticle

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