Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form

NMR structure and functional characterization of a human cancer-related nucleoside triphosphatase

Academic Article
uri icon
  • Overview
  • Research
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

related to degree

  • Placzek, William John, Ph.D. in Biology, Scripps Research 2002 - 2007

authors

  • Placzek, William John
  • Almeida, M. S.
  • Wuthrich, Kurt

publication date

  • March 2007

journal

  • Journal of Molecular Biology  Journal

abstract

  • A screen of the human cancer genome anatomy project (CGAP) database was performed to search for new proteins involved in tumorigenesis. The resulting hits were further screened for recombinant expression, solubility and protein aggregation, which led to the identification of the previously unknown human cancer-related (HCR) protein encoded by the mRNA NM_032324 as a target for structure determination by NMR. The three-dimensional structure of the protein in its complex with ATPgammaS forms a three-layered alpha/beta sandwich, with a central nine-stranded beta-sheet surrounded by five alpha-helices. Sequence and three-dimensional structure comparisons with AAA+ ATPases revealed the presence of Walker A (GPPGVGKT) and Walker B (VCVIDEIG) motifs. Using 1D (31)P-NMR spectroscopy and a coupled enzymatic assay for the determination of inorganic phosphate, we showed that the purified recombinant protein is active as a non-specific nucleoside triphosphatase, with k(cat)=7.6x10(-3) s(-1). The structural basis for the enzymatic activity of HCR-NTPase was further characterized by site-directed mutagenesis of the Walker B motif, which further contributes to making the HCR-NTPase an attractive new target for further biochemical characterization in the context of its presumed role in human tumorigenesis.

subject areas

  • Adenosine Triphosphate
  • Amino Acid Sequence
  • Humans
  • In Vitro Techniques
  • Kinetics
  • Models, Molecular
  • Molecular Sequence Data
  • Neoplasm Proteins
  • Neoplasms
  • Nuclear Magnetic Resonance, Biomolecular
  • Nucleoside-Triphosphatase
  • Protein Conformation
  • Recombinant Proteins
  • Sequence Homology, Amino Acid
scroll to property group menus

Research

keywords

  • ATP hydrolysis
  • NMR structure
  • cancer-related proteins
  • human cancers
  • nucleoside triphosphatase
scroll to property group menus

Identity

International Standard Serial Number (ISSN)

  • 0022-2836

Digital Object Identifier (DOI)

  • 10.1016/j.jmb.2007.01.001

PubMed ID

  • 17291528
scroll to property group menus

Additional Document Info

start page

  • 788

end page

  • 801

volume

  • 367

issue

  • 3

©2021 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support