Expression, purification, and circular dichroism analysis of human CDK9

Protein Expr Purif. 2006 Jun;47(2):614-20. doi: 10.1016/j.pep.2006.02.012. Epub 2006 Mar 10.

Abstract

The human cyclin-dependent kinase 9 (CDK9) protein was expressed in E. coli BL21 using the pET23a vector at 30 degrees C. Several milligrams of protein were purified from soluble fraction using ionic exchange and ATP-affinity chromatography. The structural quality of recombinant CDK9 and the estimation of its secondary structure were obtained by circular dichroism. Structural models of CDK9 presented 26% of helices in agreement with the spectra by circular dichroism analysis. This is the first report on human CDK9 expression in Escherichia coli and structure analysis and provides the first step for the development of CDK9 inhibitors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Circular Dichroism
  • Cyclin-Dependent Kinase 9 / antagonists & inhibitors
  • Cyclin-Dependent Kinase 9 / biosynthesis*
  • Cyclin-Dependent Kinase 9 / chemistry
  • Cyclin-Dependent Kinase 9 / genetics
  • Cyclin-Dependent Kinase 9 / isolation & purification*
  • Enzyme Inhibitors / chemistry
  • Escherichia coli* / genetics
  • Gene Expression
  • Humans
  • Models, Molecular
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / isolation & purification*

Substances

  • Enzyme Inhibitors
  • Recombinant Proteins
  • CDK9 protein, human
  • Cyclin-Dependent Kinase 9