Crystal structure of the kinase domain of human protein tyrosine kinase 6 (PTK6) at 2.33 Å resolution

Biochem Biophys Res Commun. 2016 Sep 16;478(2):637-42. doi: 10.1016/j.bbrc.2016.07.121. Epub 2016 Jul 30.

Abstract

Human Protein tyrosine kinase 6 (PTK6) (EC:2.7.10.2), also known as the breast tumor kinase (BRK), is an intracellular non-receptor Src-related tyrosine kinase expressed in a majority of human breast tumors and breast cancer cell lines, but its expression is low or completely absent in normal mammary glands. In the recent past, several studies have suggested that PTK6 is a potential therapeutic target in cancer. To understand its structural and functional properties, the PTK6 kinase domain (PTK6-KD) gene was cloned, overexpressed in a baculo-insect cell system, purified and crystallized at room temperature. X-ray diffraction data to 2.33 Å resolution was collected on a single PTK6-KD crystal, which belonged to the triclinic space group P1. The Matthews coefficient calculation suggested the presence of four protein molecules per asymmetric unit, with a solvent content of ∼50%.The structure has been solved by molecular replacement and crystal structure data submitted to the protein data bank under the accession number 5D7V. This is the first report of apo PTK6-KD structure crystallized in DFG-in and αC-helix-out conformation.

Keywords: BRK; Cancer; Kinase domain; Non-receptor tyrosine kinase; PTK6; X-ray crystal structure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Baculoviridae / genetics
  • Baculoviridae / metabolism
  • Cloning, Molecular
  • Crystallography, X-Ray
  • Enzyme Assays
  • Gene Expression
  • Humans
  • Kinetics
  • Models, Molecular
  • Mutation*
  • Neoplasm Proteins / chemistry*
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Protein Conformation, alpha-Helical
  • Protein Domains
  • Protein-Tyrosine Kinases / chemistry*
  • Protein-Tyrosine Kinases / genetics
  • Protein-Tyrosine Kinases / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sequence Alignment
  • Sf9 Cells
  • Spodoptera
  • Structure-Activity Relationship

Substances

  • Neoplasm Proteins
  • Recombinant Proteins
  • Protein-Tyrosine Kinases
  • PTK6 protein, human