Proton Affinities and Photoelectron Spectra of Phenylalanine and N-Methyl- and N,N-Dimethylphenylalanine. Correlation of Lone Pair Ionization Energies with Proton Affinities and Implications for N-Methylation as a Method to Effect Site Specific Protonation of Peptides
A Fourier transform ion cyclotron resonance (FT-ICR) technique for measuring gas-phase proton affinities is presented which utilizes collisional dissociation of proton-bound clusters by off-resonance translational excitation. A simplified RRKM analysis relates unimolecular dissociation rates to proton affinities. This technique is used to measure values for the proton affinities of phenylalanine and N-methyl- and N,N-dimethylphenylalanine of 220.3, 223.6, and 224.5 kcal/mol, respectively (relative to the proton affinity of NH_3 = 204.0 kcal/mol). The proton affinity measured for phenylalanine is in excellent agreement with reported literature values. The photoelectron spectra of these three molecules are also presented and analyzed. Assignments of bands to specific ionization processes are aided by comparison with model compounds such as methyl-substituted amines and 2-phenylethylamines. These data are employed to examine the correlation of adiabatic nitrogen lone pair ionization energies with gas-phase proton affinities for phenylalanine, N-methylphenylalanine, and N,N-dimethylphenylalanine in comparison to correlations for other amino acids and selected aliphatic amines. Although amine nitrogen methylation increases the potential for localizing charge at the amine terminus of protonated peptides by increasing the gas-phase proton affinity, the present study establishes that the increase is not sufficient to compete with protonation of some of the more basic side chains in peptides.
© 1994 American Chemical Society. Received October 25, 1993. Revised Manuscript Received March 31, 1994. J.L.B. gratefully acknowledges the Beckman Foundation and Institute for the initial funding and continuing support of the research facilities, and the National Science Foundation for their funding (CHE-9108318). We acknowledge the financial support of S.C. from a NIH-NRSA traineeship in Biotechnology, E.M.M. from a NIH-NRSA Human Genome traineeship and Rainin Fellowship, and M.T.R. from a California Institute of Technology Consortium grant. D.L.L. acknowledges support of the Department of Energy (Division of Chemical Sciences, Office of Basic Energy Sciences, Office of Energy Research; Contract No. DE-FG02-86ER13501), the National Science Foundation for assistance in support for instrumentation (CHE-9300841) and for support of K.F.S. under the REU program (CHE-376800), and the Materials Characterization Program (Arizona).