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Binding of guanine nucleotides to bronchial membranes: Effect of maturation

Binding of guanine nucleotides to bronchial membranes: Effect of maturation

Experimental Lung Research 24(5): 675-684

ISSN/ISBN: 0190-2148

PMID: 9779376

DOI: 10.3109/01902149809099587

Guanosine 5-[y-thio]triphosphate ([35S]GTP gamma S) binding to guinea pig bronchial membranes from immature and mature guinea pigs was rapid (Kon: 3.8 x 10(5) mol-1 min-1), saturable (Bmax: 160 pmoles/mg protein) and of high affinity (Kd: 0.6 microM). [35S]GTP gamma S rapidly dissociated in the absence of magnesium (Koff: 0.06 min-1), but 50 mM magnesium inhibited the dissociation. Maturation did not alter the affinity of the ligand, but Bmax (pmoles/mg DNA) was greater in preparations from mature animals (929 +/- 16 vs. 620 +/- 64). [35S]GTP gamma S was displaced by guanine nucleotides with a rank order of potency of GDP beta S = Gpp(NH)p > GDP > GTP, but not by ATP. We conclude that [35S]GTP gamma S is a specific and useful method to quantitate bronchial membrane-bound GTP-binding proteins. The technique shows that there is a significant increase in the cellular content of G-proteins during maturation.

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Accession: 008223629

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