Interaction studies between the p21Cip1/Waf1 cyclin-dependent kinase inhibitor and proliferating cell nuclear antigen (PCNA) by surface plasmon resonance

Martine Knibiehler, Françoise Goubin, Nathalie Escalas, Zophonias O. Jónsson, Honoré Mazarguil, Ulrich Hübscher, Bernard Ducommun*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

23 Citations (Scopus)

Abstract

The cyclin-dependent kinase (CDK) inhibitor p21Cip1 consists of two domains that interact with CDKs and proliferating cell nuclear antigen (PCNA), respectively. We have investigated the interaction between p21Cip1 and PCNA using surface plasmon resonance (SPR) technology and compared the results with those obtained from other sources such as the yeast two-hybrid system. Whilst other methods are only semiquantitative, the SPR technique allowed us to determine the kinetic parameters of the interaction. The apparent equilibrium constant K(D) calculated for these kinetic parameters was 3.2 x 10-7 M. We further demonstrate the use of SPR to study the interaction between mutant proteins and to determine their actual K(D). The interaction between p21Cip1/PCNA is shown to he dependent upon the trimeric conformation of PCNA since a point mutant that abolishes PCNA-PCNA interaction also abolishes PCNA's interaction with p21Cip1. Finally, we demonstrate that SPR can be used to characterise the interaction of p21Cip1 and PCNA in the presence of short competitive peptides.

Original languageEnglish
Pages (from-to)66-70
Number of pages5
JournalFEBS Letters
Volume391
Issue number1-2
DOIs
Publication statusPublished - 5 Aug 1996

Other keywords

  • Cell cycle
  • Cyclin-dependent kinase inhibitor
  • p21Cip1/Waf1
  • PCNA

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