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      Permanent link:
      https://www.ias.ac.in/article/fulltext/jbsc/023/03/0193-0200

    • Keywords

       

      G-proteins; ADP-ribosylation; anti-Gα (common); Funaria

    • Abstract

       

      Improved methods are described for the detection of G1P-binding proteins (G-proteins) in the protonema of mossFunaria hygrometrica and coleoptiles of corn(Zea mays) and sorghum(Sorghum vulgare). We optimized conditions for the transfer of proteins to nitrocellulose, production of high titer polyclonal anti-Gα (common) antibodies and finally the detection of G-proteins by amplification. In addition to the α-subunit of heterotrimeric G-proteins (Mr 41–43 kDa), a small molecular weight class (< 30 kDa) was also detected by anti-Gα (common) antibodies. An easy, reliable and efficient filter assay is also described to quantify the toxin catalyzed ADP-ribosylation. The apparentKm of the NAD has been determined to be approximately 1.5μM for the microsomal fraction of moss. Inclusion of G1P stimulated ADP-ribosylation by 2–27-fold. One to three polypeptides representing the α-subunit of heterotrimeric G-proteins of (Mr 37–43 kDa) were ADP-ribosylated in all three plants. The anti-Gβ (C-terminus) antibody cross-reacted strongly with 39 and 34 kDa polypeptide in moss and corn respectively. By employing improved methods two classes of G-proteins have been shown to be present in three plant species.

    • Author Affiliations

       

      Kishore C S Panigrahi1 M M Johri1

      1. Department of Biological Sciences, Tata Institute of Fundamental Research, Homi Bhabha Road, Mumbai - 400005, India
    • Dates

       
  • Journal of Biosciences | News

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