1mx4 › Citations

Structure-based design of p18INK4c proteins with increased thermodynamic stability and cell cycle inhibitory activity.

Venkataramani RN, MacLachlan TK, Chai X, El-Deiry WS, Marmorstein R
J Biol Chem 277 48827-33 (2002)
Related entries: 1mx2, 1mx6

Cited: 9 times
EuropePMC logo PMID: 12370184

Abstract

p18(INK4c) is a member of the INK4 family of proteins that regulate the G(1) to S cell cycle transition by binding to and inhibiting the pRb kinase activity of cyclin-dependent kinases 4 and 6. The p16(INK4a) member of the INK4 protein family is altered in a variety of cancers and structure-function studies of the INK4 proteins reveal that the vast majority of missense tumor-derived p16(INK4a) mutations reduce protein thermodynamic stability. Based on this observation, we used p18(INK4c) as a model to test the proposal that INK4 proteins with increased stability might have enhanced cell cycle inhibitory activity. Structure-based mutagenesis was used to prepare p18(INK4c) mutant proteins with a predicted increase in stability. Using this approach, we report the generation of three mutant p18(INK4C) proteins, F71N, F82Q, and F92N, with increased stability toward thermal denaturation of which the F71N mutant also showed an increased stability to chemical denaturation. The x-ray crystal structures of the F71N, F82Q, and F92N p18INK4C mutant proteins were determined to reveal the structural basis for their increased stability properties. Significantly, the F71N mutant also showed enhanced CDK6 interaction and cell cycle inhibitory activity in vivo, as measured using co-immunoprecipitation and transient transfection assays, respectively. These studies show that a structure-based approach to increase the thermodynamic stability of INK4 proteins can be exploited to prepare more biologically active molecules with potential applications for the development of molecules to treat p16(INK4a)-mediated cancers.

Reviews - 1mx4 mentioned but not cited (1)

  1. A primer on ankyrin repeat function in TRP channels and beyond. Gaudet R. Mol Biosyst 4 372-379 (2008)


Reviews citing this publication (1)

  1. Selectivity and potency of cyclin-dependent kinase inhibitors. Sridhar J, Akula N, Pattabiraman N. AAPS J 8 E204-21 (2006)

Articles citing this publication (7)

  1. The energy landscapes of repeat-containing proteins: topology, cooperativity, and the folding funnels of one-dimensional architectures. Ferreiro DU, Walczak AM, Komives EA, Wolynes PG. PLoS Comput Biol 4 e1000070 (2008)
  2. P18 is a tumor suppressor gene involved in human medullary thyroid carcinoma and pheochromocytoma development. van Veelen W, Klompmaker R, Gloerich M, van Gasteren CJ, Kalkhoven E, Berger R, Lips CJ, Medema RH, Höppener JW, Acton DS. Int J Cancer 124 339-345 (2009)
  3. Small-molecule inhibitors targeting INK4 protein p18(INK4C) enhance ex vivo expansion of haematopoietic stem cells. Gao Y, Yang P, Shen H, Yu H, Song X, Zhang L, Zhang P, Cheng H, Xie Z, Hao S, Dong F, Ma S, Ji Q, Bartlow P, Ding Y, Wang L, Liu H, Li Y, Cheng H, Miao W, Yuan W, Yuan Y, Cheng T, Xie XQ. Nat Commun 6 6328 (2015)
  4. Differential post-transcriptional regulation of two Ink4 proteins, p18 Ink4c and p19 Ink4d. Forget A, Ayrault O, den Besten W, Kuo ML, Sherr CJ, Roussel MF. Cell Cycle 7 3737-3746 (2008)
  5. Discovery of novel INK4C small-molecule inhibitors to promote human and murine hematopoietic stem cell ex vivo expansion. Xie XQ, Yang P, Zhang Y, Zhang P, Wang L, Ding Y, Yang M, Tong Q, Cheng H, Ji Q, McGuire T, Yuan W, Cheng T, Gao Y. Sci Rep 5 18115 (2015)
  6. A Conserved Gammaherpesvirus Cyclin Specifically Bypasses Host p18(INK4c) To Promote Reactivation from Latency. Williams LM, Niemeyer BF, Franklin DS, Clambey ET, van Dyk LF. J Virol 89 10821-10831 (2015)
  7. Cyclin dependent kinase inhibitors differentially modulate synergistic cytokine responsiveness of hematopoietic progenitor cells. Broxmeyer HE, Franklin DS, Cooper S, Hangoc G, Mantel C. Stem Cells Dev 21 1597-1603 (2012)


Quick links