+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Regulatory GTP-binding proteins (ADP-ribosylation factor, Gt, and RAS) are not activated directly by nucleoside diphosphate kinase



Regulatory GTP-binding proteins (ADP-ribosylation factor, Gt, and RAS) are not activated directly by nucleoside diphosphate kinase



Journal of Biological Chemistry 267(25): 18182



The expression of nucleoside diphosphate kinase (NDK) genes has been implicated as a negative regulator of murine and human tumor metastases and is critical to proper development in Drosophila melanogaster. Molecular mechanisms for the role(s) of NDK in these complex processes have not yet been elucidated, but several reports have suggested that these and many other signal transduction pathways may be activated by NDK acting directly on a regulatory GTP-binding protein(s). To test this hypothesis, we examined the ability of NDK to catalyze the phosphorylation of the GDP bound to the following three members of the superfamily of regulatory GTP-binding proteins: Gt, Ha-ras p21, and ARF. We have found no evidence to support the hypothesis that NDK can directly activate any GTP-binding protein. Rather, evidence is presented which clearly shows that all of the GTP formed upon incubation of GTP-binding proteins with NDK is the result of NDK utilizing free GDP as substrate. The GDP bound to the regulatory proteins is not a substrate for NDK under conditions in which free nucleotides are rapidly and efficiently phosphorylated. The importance of appropriate controls for dissociation of GDP from the regulatory proteins both during the NDK reaction and during the analysis of product is demonstrated. We believe there is currently no experimental evidence to support the hypothesis that NDK can directly activate a regulatory GTP-binding protein.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 041214053

Download citation: RISBibTeXText

PMID: 1325460


Related references

Microtubules and nucleoside diphosphate kinase nucleoside diphosphate kinase binds to co purifying contaminants rather than microtubule proteins. Biochemical Journal 232(3): 651-656, 1985

Role of binding and nucleoside diphosphate kinase A in the regulation of the cystic fibrosis transmembrane conductance regulator by AMP-activated protein kinase. Journal of Biological Chemistry 287(40): 33389-33400, 2012

Conversion of GDP into GTP by nucleoside diphosphate kinase on the GTP-binding proteins. Journal of Biological Chemistry 265(35): 21536-21540, 1990

Nucleoside diphosphate kinase/nm23 as a key regulatory factor of tumor metastasis and cell motility. Biologicheskie Membrany (Moscow) 20(1): 59-65, January-February, 2003

Nucleoside diphosphate kinase and GTP-binding proteins. Possible mechanisms of coupling. Biofizika 53(6): 922-928, 2008

Escherichia coli nucleoside diphosphate kinase interactions with T4 phage proteins of deoxyribonucleotide synthesis and possible regulatory functions. Journal of Biological Chemistry 279(31): 32225-32232, 2004

Physiological correlation between nucleoside-diphosphate kinase and the enzyme-associated guanine nucleotide binding proteins. Biochemical and Biophysical Research Communications 143(2): 552-559, 1987

Nucleoside diphosphate kinase does not directly interact with tubulin nor microtubules. Biochemical and Biophysical Research Communications 187(1): 65-72, 1992

Direct activation of guanine nucleotide binding proteins through a high-energy phosphate-transfer by nucleoside diphosphate-kinase. Biochemical and Biophysical Research Communications 148(1): 300-307, 1987

X-ray analysis of azido-thymidine diphosphate binding to nucleoside diphosphate kinase. Proceedings of the National Academy of Sciences of the United States of America 94(14): 7162-7165, 1997

Adenosine 5'-diphosphate binding and the active site of nucleoside diphosphate kinase. Biochemistry 33(2): 459-467, 1994

Nucleoside diphosphate kinase of Mycobacterium smegmatis Identification of proteins that modulate specificity of nucleoside triphosphate synthesis by the enzyme. Abstracts of the General Meeting of the American Society for Microbiology 97: 567, 1997