Identification and localization of pertussis toxin-sensitive GTP-binding proteins in bovine kidney glomeruli

Ajay Gupta, Bahar Bastani, Henry Purcell, Keith A. Hruska

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2 Scopus citations


The vascular tree and the mesangium in the glomerulus respond to various hormones, growth factors, and autonomic signals, leading to generation of second messengers and regulation of ion channels. Guanine nucleotide regulatory proteins (G proteins) mediate these effects in other systems. Glomerular G proteins were studied by immunoblotting and immunohistochemical techniques. Glomeruli were isolated from bovine kidney cortex by differential sieving. Glomerular proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and nitrocellulose transfers were immunoblotted with antibodies to G proteins. Gα,Common antiserum (P-960) recognized proteins with a molecular mass of 41 to 45 kDa. Antibodies against peptide sequences specific to G and G demonstrated Giα,1/3 (molecular mass, 39 to 41 kDa), Giα2 (molecular mass, 40 kDa), and G (molecular mass, 39 kDa). Presence of these proteins was further confirmed by pertussis toxin-catalyzed ADP ribosylation of protein(s) with a molecular mass of 39 to 41 kDa in the glomeruli. Immunohistochemical staining of frozen sections from bovine kidney cortex revealed the presence of Giα2 in capillary loop distribution in glomeruli and interstitium, but Gi,1/3 or Go could not be demonstrated. The pattern of immunofluorescence with Giα2 antiserum suggested localization of Giα2 to the endothelium in glomerular and interstitial vasculature. The novel finding of Go in glomeruli requires localization of Go to specific cells and determination of its role in glomerular physiology. In conclusion, these studies demonstrate that bovine kidney glomeruli express a subunits of pertussis toxin-sensitive GTP-binding proteins Gi,1/3, Gi,2 and Go. Giα2 is localized to the vascular endothelium in glomeruli and interstitium, but Gi,1/3 could not be localized. Further work is required to determine the role of these proteins in mediating effects of neuronal stimuli, hormones, and cytokines on glomerular blood flow.

Original languageEnglish
Pages (from-to)172-178
Number of pages7
JournalJournal of the American Society of Nephrology
Issue number2
StatePublished - Aug 1991


  • ADP ribosylation
  • Endothelium
  • G
  • Immunoblotting
  • Immunohistochemistry


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