The crystal structure of human protein farnesyltransferase reveals the basis for inhibition by CaaX tetrapeptides and their mimetics
10.1073/pnas.241407898
Crossref journal-article
Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences (341)
Abstract

Protein farnesyltransferase (FTase) catalyzes the attachment of a farnesyl lipid group to the cysteine residue located in the C-terminal tetrapeptide of many essential signal transduction proteins, including members of the Ras superfamily. Farnesylation is essential both for normal functioning of these proteins, and for the transforming activity of oncogenic mutants. Consequently FTase is an important target for anti-cancer therapeutics. Several FTase inhibitors are currently undergoing clinical trials for cancer treatment. Here, we present the crystal structure of human FTase, as well as ternary complexes with the TKCVFM hexapeptide substrate, CVFM non-substrate tetrapeptide, and L-739,750 peptidomimetic with either farnesyl diphosphate (FPP), or a nonreactive analogue. These structures reveal the structural mechanism of FTase inhibition. Some CaaX tetrapeptide inhibitors are not farnesylated, and are more effective inhibitors than farnesylated CaaX tetrapeptides. CVFM and L-739,750 are not farnesylated, because these inhibitors bind in a conformation that is distinct from the TKCVFM hexapeptide substrate. This non-substrate binding mode is stabilized by an ion pair between the peptide N terminus and the α-phosphate of the FPP substrate. Conformational mapping calculations reveal the basis for the sequence specificity in the third position of the CaaX motif that determines whether a tetrapeptide is a substrate or non-substrate. The presence of β-branched amino acids in this position prevents formation of the non-substrate conformation; all other aliphatic amino acids in this position are predicted to form the non-substrate conformation, provided their N terminus is available to bind to the FPP α-phosphate. These results may facilitate further development of FTase inhibitors.

Bibliography

Long, S. B., Hancock, P. J., Kral, A. M., Hellinga, H. W., & Beese, L. S. (2001). The crystal structure of human protein farnesyltransferase reveals the basis for inhibition by CaaX tetrapeptides and their mimetics. Proceedings of the National Academy of Sciences, 98(23), 12948–12953.

Authors 5
  1. Stephen B. Long (first)
  2. Paula J. Hancock (additional)
  3. Astrid M. Kral (additional)
  4. Homme W. Hellinga (additional)
  5. Lorena S. Beese (additional)
References 37 Referenced 92
  1. P J Casey Science 268, 221–225 (1995). (10.1126/science.7716512) / Science by Casey P J (1995)
  2. F L Zhang, P J Casey Annu Rev Biochem 65, 241–269 (1996). (10.1146/annurev.bi.65.070196.001325) / Annu Rev Biochem by Zhang F L (1996)
  3. H R Ashar, L James, K Gray, D Carr, S Black, L Armstrong, W R Bishop, P Kirschmeier J Biol Chem 275, 30451–30457 (2000). (10.1074/jbc.M003469200) / J Biol Chem by Ashar H R (2000)
  4. W R Schafer, R Kim, R Sterne, J Thorner, S-H Kim, J Rine Science 245, 379–385 (1989). (10.1126/science.2569235) / Science by Schafer W R (1989)
  5. P J Casey, P A Solski, C J Der, J E Buss Proc Natl Acad Sci USA 86, 8323–8327 (1989). (10.1073/pnas.86.21.8323) / Proc Natl Acad Sci USA by Casey P J (1989)
  6. J F Hancock, A I Magee, J E Childs, C J Marshall Cell 57, 1167–1177 (1989). (10.1016/0092-8674(89)90054-8) / Cell by Hancock J F (1989)
  7. Y Reiss, J L Goldstein, M C Seabra, P J Casey, M S Brown Cell 62, 81–88 (1990). (10.1016/0092-8674(90)90242-7) / Cell by Reiss Y (1990)
  8. J L Goldstein, M S Brown, S J Stradley, Y Reiss, L M Geirasch J Biol Chem 266, 15575–15578 (1991). (10.1016/S0021-9258(18)98441-3) / J Biol Chem by Goldstein J L (1991)
  9. M S Brown, J L Goldstein, K J Paris, J P Burnier, J C Marsters Proc Natl Acad Sci USA 89, 8313–8316 (1992). (10.1073/pnas.89.17.8313) / Proc Natl Acad Sci USA by Brown M S (1992)
  10. S L Moores, M D Schaber, S D Mosser, E Rands, M B O'Hara, V M Garsky, M S Marshall, D L Pompliano, J B Gibbs J Biol Chem 266, 14603–14610 (1991). (10.1016/S0021-9258(18)98729-6) / J Biol Chem by Moores S L (1991)
  11. T M Williams, C J Dinsmore Adv Med Chem 4, 273–314 (1999). (10.1016/S1067-5698(99)80008-X) / Adv Med Chem by Williams T M (1999)
  12. L Sepp-Lorenzino, Z Ma, E Rands, N E Kohl, J B Gibbs, A Oliff, N Rosen Cancer Res 55, 5302–5309 (1995). / Cancer Res by Sepp-Lorenzino L (1995)
  13. N E Kohl, C A Omer, M W Conner, N J Anthony, J P Davide, S J deSolms, E A Giuliani, R P Gomez, S L Graham, K Hamilton, et al. Nat Med 1, 792–797 (1995). (10.1038/nm0895-792) / Nat Med by Kohl N E (1995)
  14. S R D Johnston Lancet Oncol 2, 18–26 (2001). (10.1016/S1470-2045(00)00191-1) / Lancet Oncol by Johnston S R D (2001)
  15. N E Kohl, F R Wilson, S D Mosser, E Giuliani, S J deSolms, M W Conner, N J Anthony, W J Holtz, R P Gomez, T J Lee, et al. Proc Natl Acad Sci USA 91, 9141–9145 (1994). (10.1073/pnas.91.19.9141) / Proc Natl Acad Sci USA by Kohl N E (1994)
  16. K S Koblan, J C Culberson, S J deSolms, E A Giuliani, S D Mosser, C A Omer, S M Pitzenberger, M J Bogusky Protein Sci 4, 681–688 (1995). (10.1002/pro.5560040408) / Protein Sci by Koblan K S (1995)
  17. C L Strickland, W T Windsor, R Syto, L Wang, R Bond, Z Wu, J Schwartz, H V Le, L S Beese, P C Weber Biochemistry 37, 16601–16611 (1998). (10.1021/bi981197z) / Biochemistry by Strickland C L (1998)
  18. S B Long, P J Casey, L S Beese Structure Fold Des 8, 209–222 (2000). (10.1016/S0969-2126(00)00096-4) / Structure Fold Des by Long S B (2000)
  19. C-C Huang, P J Casey, C A Fierke J Biol Chem 272, 20–23 (1997). (10.1074/jbc.272.1.20) / J Biol Chem by Huang C-C (1997)
  20. K E Hightower, C-C Huang, P J Casey, C A Fierke Biochemistry 37, 15555–15562 (1998). (10.1021/bi981525v) / Biochemistry by Hightower K E (1998)
  21. S P Eisenberg, R J Evans, W P Arend, E Verderber, M T Brewer, C H Hannum, R C Thompson Nature (London) 343, 341–346 (1990). (10.1038/343341a0) / Nature (London) by Eisenberg S P (1990)
  22. C A Omer, A M Kral, R E Diehl, G C Prendergast, S Powers, C M Allen, J B Gibbs, N E Kohl Biochemistry 32, 5167–5176 (1993). (10.1021/bi00070a028) / Biochemistry by Omer C A (1993)
  23. C A Omer, R E Diehl, A M Kral Methods Enzymol 250, 3–12 (1995). (10.1016/0076-6879(95)50057-X) / Methods Enzymol by Omer C A (1995)
  24. H-W Park, S R Boduluri, J F Moomaw, P J Casey, L S Beese Science 275, 1800–1804 (1997). (10.1126/science.275.5307.1800) / Science by Park H-W (1997)
  25. W-J Chen, J F Moomaw, L Overton, T A Kost, P J Casey J Biol Chem 268, 9675–9680 (1993). (10.1016/S0021-9258(18)98402-4) / J Biol Chem by Chen W-J (1993)
  26. V Manne, C S Ricca, J G Brown, A V Tuomari, N Yan, D Patel, R Schmidt, M J Lynch, C P Ciosek, J M Carboni, et al. Drug Dev Res 34, 121–137 (1995). (10.1002/ddr.430340205) / Drug Dev Res by Manne V (1995)
  27. C L Strickland, P C Weber, W T Windsor, Z Wu, H V Le, M M Albanese, C S Alvarez, D Cesarz, J del Rosario, J Deskus, et al. J Med Chem 42, 2125–2135 (1999). (10.1021/jm990030g) / J Med Chem by Strickland C L (1999)
  28. Z Otwinowski, W Minor Methods Enzymol 276A, 307–326 (1997). (10.1016/S0076-6879(97)76066-X) / Methods Enzymol by Otwinowski Z (1997)
  29. A T Brünger x-plor, A System for X-Ray Crystallography and NMR (Yale Univ. Press, New Haven, CT, , Version 3.1. (1992). / x-plor, A System for X-Ray Crystallography and NMR by Brünger A T (1992)
  30. A T Brünger, J S Jiang J Mol Biol 243, 100–115 (1994). (10.1006/jmbi.1994.1633) / J Mol Biol by Brünger A T (1994)
  31. S Long, P J Casey, L S Beese Biochemistry 37, 9612–9618 (1998). (10.1021/bi980708e) / Biochemistry by Long S (1998)
  32. S J Stradley, J Rizo, L M Gierasch Biochemistry 32, 12586–12590 (1993). (10.1021/bi00210a006) / Biochemistry by Stradley S J (1993)
  33. N E Kohl, S D Mosser, S J deSolms, E A Giuliani, D L Pompliano, S L Graham, R L Smith, E M Scolnick, A Oliff, J B Gibbs Science 260, 1934–1942 (1993). (10.1126/science.8316833) / Science by Kohl N E (1993)
  34. J M Word, S C Lovell, T H LaBean, H C Taylor, M E Zalis, B K Presley, J S Richardson, D C Richardson J Mol Biol 285, 1711–1733 (1999). (10.1006/jmbi.1998.2400) / J Mol Biol by Word J M (1999)
  35. T A Jones, M Kjeldgaard o, The Manual (Uppsala University, Uppsala, , Version 5.9. (1993). / o, The Manual by Jones T A (1993)
  36. H W Hellinga, F M Richards J Mol Biol 222, 763–785 (1991). (10.1016/0022-2836(91)90510-D) / J Mol Biol by Hellinga H W (1991)
  37. L L Looger, H W Hellinga J Mol Biol 307, 429–445 (2001). (10.1006/jmbi.2000.4424) / J Mol Biol by Looger L L (2001)
Dates
Type When
Created 23 years, 1 month ago (July 26, 2002, 10:31 a.m.)
Deposited 3 years, 2 months ago (June 7, 2022, 1:13 a.m.)
Indexed 1 week ago (Aug. 30, 2025, 1:12 p.m.)
Issued 23 years, 10 months ago (Oct. 30, 2001)
Published 23 years, 10 months ago (Oct. 30, 2001)
Published Online 23 years, 10 months ago (Oct. 30, 2001)
Published Print 23 years, 10 months ago (Nov. 6, 2001)
Funders 0

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@article{Long_2001, title={The crystal structure of human protein farnesyltransferase reveals the basis for inhibition by CaaX tetrapeptides and their mimetics}, volume={98}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.241407898}, DOI={10.1073/pnas.241407898}, number={23}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Long, Stephen B. and Hancock, Paula J. and Kral, Astrid M. and Hellinga, Homme W. and Beese, Lorena S.}, year={2001}, month=oct, pages={12948–12953} }