H-Ras peptide and protein substrates bind protein farnesyltransferase as an ionized thiolate - PubMed (original) (raw)
. 1998 Nov 3;37(44):15555-62.
doi: 10.1021/bi981525v.
Affiliations
- PMID: 9799520
- DOI: 10.1021/bi981525v
H-Ras peptide and protein substrates bind protein farnesyltransferase as an ionized thiolate
K E Hightower et al. Biochemistry. 1998.
Abstract
The zinc metalloenzyme protein farnesyltransferase (FTase) catalyzes the alkylation of a cysteine residue of protein substrates with a 15 carbon farnesyl group. We have developed fluorescence assays to directly measure the affinity of the enzyme for peptide and protein (Ras) substrates. A peptide corresponding to the carboxyl terminus of H-Ras binds to FTase in the microM range (KD = 4 microM) at physiological pH; however, the peptide affinity is enhanced approximately 70-fold in a ternary complex with an enzyme-bound farnesyl diphosphate (FPP) analogue, indicating that the two substrates bind synergistically. The pH dependence of substrate binding was also investigated, and two ionizations were observed: for the ternary complex, the pKa values are 8.1, reflecting ionization of the thiol of the free peptide, and 6.4. The pH dependence of the ligand-metal charge-transfer band in the optical absorption spectra of a Co2+-substituted FTase ternary complex suggests that a metal-coordinated thiol ionizes with a pKa of 6.3. These data indicate that metal coordination of the peptide sulfur with the zinc ion in FTase lowers the pKa of the thiol resulting in formation of a bound thiolate at physiological pH.
Similar articles
- Farnesylation of nonpeptidic thiol compounds by protein farnesyltransferase.
Hightower KE, Casey PJ, Fierke CA. Hightower KE, et al. Biochemistry. 2001 Jan 30;40(4):1002-10. doi: 10.1021/bi002237d. Biochemistry. 2001. PMID: 11170422 - Modulation of the zinc(II) center in protein farnesyltransferase by mutagenesis of the zinc(II) ligands.
Harris CM, Derdowski AM, Poulter CD. Harris CM, et al. Biochemistry. 2002 Aug 20;41(33):10554-62. doi: 10.1021/bi020349u. Biochemistry. 2002. PMID: 12173942 - Protein farnesyltransferase.
Park HW, Beese LS. Park HW, et al. Curr Opin Struct Biol. 1997 Dec;7(6):873-80. doi: 10.1016/s0959-440x(97)80160-1. Curr Opin Struct Biol. 1997. PMID: 9434909 Review. - Zinc-catalyzed sulfur alkyation:insights from protein farnesyltransferase.
Hightower KE, Fierke CA. Hightower KE, et al. Curr Opin Chem Biol. 1999 Apr;3(2):176-81. doi: 10.1016/s1367-5931(99)80030-1. Curr Opin Chem Biol. 1999. PMID: 10226042 Review.
Cited by
- From quantum-derived principles underlying cysteine reactivity to combating the COVID-19 pandemic.
Mazmanian K, Chen T, Sargsyan K, Lim C. Mazmanian K, et al. Wiley Interdiscip Rev Comput Mol Sci. 2022 Sep-Oct;12(5):e1607. doi: 10.1002/wcms.1607. Epub 2022 Mar 5. Wiley Interdiscip Rev Comput Mol Sci. 2022. PMID: 35600063 Free PMC article. - Sequence-Selective Covalent CaaX-Box Receptors Prevent Farnesylation of Oncogenic Ras Proteins and Impact MAPK/PI3 K Signaling.
Franz M, Mörchen B, Degenhart C, Gülden D, Shkura O, Wolters D, Koch U, Klebl B, Stoll R, Helfrich I, Scherkenbeck J. Franz M, et al. ChemMedChem. 2021 Aug 19;16(16):2504-2514. doi: 10.1002/cmdc.202100167. Epub 2021 May 19. ChemMedChem. 2021. PMID: 33899342 Free PMC article. - SmgGDS-607 Regulation of RhoA GTPase Prenylation Is Nucleotide-Dependent.
Jennings BC, Lawton AJ, Rizk Z, Fierke CA. Jennings BC, et al. Biochemistry. 2018 Jul 24;57(29):4289-4298. doi: 10.1021/acs.biochem.8b00567. Epub 2018 Jul 10. Biochemistry. 2018. PMID: 29940100 Free PMC article. - Mechanistic Understanding of Lanthipeptide Biosynthetic Enzymes.
Repka LM, Chekan JR, Nair SK, van der Donk WA. Repka LM, et al. Chem Rev. 2017 Apr 26;117(8):5457-5520. doi: 10.1021/acs.chemrev.6b00591. Epub 2017 Jan 30. Chem Rev. 2017. PMID: 28135077 Free PMC article. Review. - Role of substrate dynamics in protein prenylation reactions.
Chakravorty DK, Merz KM Jr. Chakravorty DK, et al. Acc Chem Res. 2015 Feb 17;48(2):439-48. doi: 10.1021/ar500321u. Epub 2014 Dec 24. Acc Chem Res. 2015. PMID: 25539152 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials
Miscellaneous