Purification and crystallization of cyclin-dependent kinase inhibitor p21

Protein Sci. 1996 Sep;5(9):1928-30. doi: 10.1002/pro.5560050920.

Abstract

p21, a universal inhibitor of mammalian cyclin-dependent kinases (CDK), regulates cell cycle progression by forming various distinct protein complexes with cyclins, CDKs, and the proliferating cell nuclear antigen. We have overexpressed recombinant human p21 in E. coli and purified active p21 to near homogeneity on a large scale. Crystals of recombinant p21 have been grown in the space group P2(1) a = 157.4, b = 152.7, c = 90.6 A, and beta = 92.7 degrees. The diffraction data of the recombinant p21 have been collected to 2.5 and 3.5 A resolution for the native crystal and two heavy atom derivatives of mercury and iridium.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Crystallization
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins / chemistry*
  • Cyclins / isolation & purification*
  • Enzyme Inhibitors* / chemistry
  • Enzyme Inhibitors* / isolation & purification
  • Escherichia coli / genetics
  • Gene Expression
  • Humans
  • Iridium
  • Mercury
  • Proliferating Cell Nuclear Antigen / pharmacology
  • Recombinant Proteins / chemistry
  • X-Ray Diffraction

Substances

  • CDKN1A protein, human
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins
  • Enzyme Inhibitors
  • Proliferating Cell Nuclear Antigen
  • Recombinant Proteins
  • Iridium
  • Mercury