A nucleoside diphosphate kinase A (nm23-H1) serine 120-->glycine substitution in advanced stage neuroblastoma affects enzyme stability and alters protein-protein interaction.

Abstract

A high level of nucleoside diphosphate kinase A (NDPK A/nm23-H1) in neuroblastoma is associated with advanced stage disease. We have also found a serine 120-->glycine substitution in NDPK A and/or amplification of the nm23-H1 gene in advanced stage neuroblastomas. Serine 120, a highly conserved residue, is located in proximity to histidine 118 which forms a phosphorylated intermediate essential for NDPK activity. The effect of Ser120-->Gly substitution on the biochemical properties of NDPK A was investigated. Phosphate-transferase activity was lower in the recombinant mutant NDPK A and in the immunoprecipitated complex consisting of NDPK A and NDPK B prepared from a neuroblastoma tumor containing the mutation, relative to the wild-type. There was a significant decrease in the enzyme stability toward urea- or temperature-induced denaturation for the recombinant mutant NDPK A and in an immunoprecipitate from a tumor containing the mutation. Recombinant NDPK A containing the Ser120-->Gly mutation exhibited reduced hexameric and increased dimeric oligomerization relative to the wild-type. Moreover a 28 kDa cellular protein was detected, that co-precipitated with the mutant but not wild-type NDPK A. The altered properties of the mutant protein may have relevance to a role for NDPK A in neuroblastoma progression.

• Authors: C L Chang, J R Strahler, D H Thoraval, M G Qian, R Hinderer, S M Hanash
• Journal: Oncogene (Oncogene) Vol. 12 Issue 3 Pg. 659-67 (Feb 01 1996) ISSN: 0950-9232 [Print] England
• PMID: 8637723 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Cross-Linking Reagents
• DNA Primers
• Isoenzymes
• NM23 Nucleoside Diphosphate Kinases
• Recombinant Proteins
• Transcription Factors
• Serine
• NME1 protein, human
• Nucleoside-Diphosphate Kinase
• Monomeric GTP-Binding Proteins
• Glutaral
• Glycine

Topics

• Base Sequence
• Cross-Linking Reagents
• DNA Primers
• Enzyme Stability
• Glutaral
• Glycine
• Hot Temperature
• Humans
• Isoenzymes (chemistry, isolation & purification, metabolism)
• Kinetics
• Molecular Sequence Data
• Monomeric GTP-Binding Proteins
• Mutagenesis, Site-Directed
• NM23 Nucleoside Diphosphate Kinases
• Neoplasm Staging
• Neuroblastoma (enzymology, genetics, pathology)
• Nucleoside-Diphosphate Kinase (chemistry, metabolism)
• Point Mutation
• Polymerase Chain Reaction
• Protein Denaturation
• Recombinant Proteins (chemistry, isolation & purification, metabolism)
• Serine
• Thermodynamics
• Transcription Factors (chemistry, isolation & purification, metabolism)