PMID: 2111167Jan 30, 1990Paper

Mechanism of activation of cholera toxin by ADP-ribosylation factor (ARF): both low- and high-affinity interactions of ARF with guanine nucleotides promote toxin activation

Biochemistry
D A BobakJ Moss

Abstract

Activation of adenylyl cyclase by cholera toxin A subunit (CT-A) results from the ADP-ribosylation of the stimulatory guanine nucleotide binding protein (GS alpha). This process requires GTP and an endogenous guanine nucleotide binding protein known as ADP-ribosylation factor (ARF). One membrane (mARF) and two soluble forms (sARF I and sARF II) of ARF have been purified from bovine brain. Because the conditions reported to enhance the binding of guanine nucleotides by ARF differ from those observed to promote optimal activity, we sought to characterize the determinants influencing the functional interaction of guanine nucleotides with ARF. High-affinity GTP binding by sARF II (apparent KD of approximately 70 nM) required Mg2+, DMPC, and sodium cholate. sARF II, in DMPC/cholate, also enhanced CT-A ADP-ribosyltransferase activity (apparent EC50 for GTP of approximately 50 nM), although there was a delay before achievement of a maximal rate of sARF II stimulated toxin activity. The delay was abolished by incubation of sARF II with GTP at 30 degrees C before initiation of the assay. In contrast, a maximal rate of activation of toxin by sARF II, in 0.003% SDS, occurred without delay (apparent EC50 for GTP of approximately 5 microM)....Continue Reading

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Citations

Dec 11, 1996·Biochimica Et Biophysica Acta·O SaitohH Nakata
Nov 22, 1994·Proceedings of the National Academy of Sciences of the United States of America·D MassenburgM Vaughan
Mar 28, 1995·Proceedings of the National Academy of Sciences of the United States of America·J X HongM Vaughan
Dec 1, 1992·Molecular Biology of the Cell·M Bomsel, K Mostov
Aug 30, 2013·Nihon saikingaku zasshi. Japanese journal of bacteriology·Masatoshi Noda
Jan 10, 1998·The Journal of Biological Chemistry·F J LeeM Vaughan
Jul 16, 2014·Nature Reviews. Microbiology·Nathan C SimonJoseph T Barbieri
Feb 28, 2014·The Journal of Biological Chemistry·Nathan C Simon, Joseph T Barbieri
Apr 18, 2001·The Journal of Biological Chemistry·Z GombosM Ikura
Nov 17, 2005·DNA Sequence : the Journal of DNA Sequencing and Mapping·Lei WangYony-Jun Wang
Aug 1, 1995·Infection and Immunity·D A KnightJ T Barbieri

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