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A point mutation of human nucleoside diphosphate kinase A found in aggressive neuroblastoma affects protein folding.

Abstract
The point mutation serine 120 to glycine in the human nucleoside diphosphate kinase A has been identified in several aggressive neuroblastomas (Chang, C. L., Zhu, X. X., Thoraval, D. H., Ungar, D., Rawwas, J., Hora, N., Strahler, J. R., Hanash, S. M. & Radany, E. (1994) Nature 370, 335-336). We expressed in bacteria and purified wild-type and S120G mutant nucleoside diphosphate kinase A. The mutant enzyme had enzymatic and structural properties similar to the wild-type enzyme, whereas its stability to denaturation by heat and urea was markedly reduced. More importantly, upon renaturation of the urea-denatured mutant protein, a folding intermediate accumulated, having the characteristics of a molten globule. It had no tertiary structure, as shown by near UV circular dichroism, whereas the secondary structure was substantially recovered. The hydrophobic probe 8-anilino-1-naphthalene sulfonate bound to the intermediate species with an increase in fluorescence intensity and a blue shift. The hydrodynamic size was between that expected for a folded and an unfolded monomer. Finally, electrophoresis in a transverse urea gradient displayed no renaturation curve, and the protein showed the tendency to aggregate at the lowest urea concentrations. The existence of a molten globule folding intermediates resulting from an altered folding in the mutated protein might be related to the aggressiveness of neuroblastomas.
AuthorsI Lascu, S Schaertl, C Wang, C Sarger, A Giartosio, G Briand, M L Lacombe, M Konrad
JournalThe Journal of biological chemistry (J Biol Chem) Vol. 272 Issue 25 Pg. 15599-602 (Jun 20 1997) ISSN: 0021-9258 [Print] United States
PMID9188446 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Antigens, Neoplasm
  • NM23 Nucleoside Diphosphate Kinases
  • Transcription Factors
  • Serine
  • Nucleoside-Diphosphate Kinase
  • Monomeric GTP-Binding Proteins
  • Glycine
Topics
  • Antigens, Neoplasm (chemistry, genetics)
  • Chromatography, Gel
  • Circular Dichroism
  • Glycine
  • Humans
  • Kinetics
  • Molecular Weight
  • Monomeric GTP-Binding Proteins
  • NM23 Nucleoside Diphosphate Kinases
  • Neuroblastoma (enzymology, genetics)
  • Nucleoside-Diphosphate Kinase (chemistry, genetics)
  • Point Mutation
  • Protein Folding
  • Serine
  • Transcription Factors (chemistry, genetics)

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