Spectroscopic Characterization of the SH2- and Active Site-Directed Peptide Sequences of a Bivalent Src Kinase Inhibitor

• Published in: Applied spectroscopy Vol. 63; no. 7; pp. 767 - 774
• Main Authors: Desamero, Ruel Z. B., Kang, Jeonghee, Dol, Chrystel, Chinwong, Justina, Walters, Karim, Sivarajah, Thulashie, Profit, Adam A.
• Format: Journal Article
• Language: English
• Published: London, England SAGE Publications 01-07-2009
• Subjects: Catalytic Domain; Computer Simulation; Models, Chemical; Peptides - chemistry; Peptides - pharmacology; Phenylalanine - chemistry; Phosphotyrosine - chemistry; Protein Kinase Inhibitors - chemistry; Protein Kinase Inhibitors - pharmacology; Solvents - chemistry; Spectroscopy, Fourier Transform Infrared - methods; src Homology Domains; src-Family Kinases - antagonists & inhibitors; src-Family Kinases - chemistry; src-Family Kinases - metabolism; FT-IR spectroscopy; DFT; Phosphotyrosine; Pentafluorophenylalanine; Bivalent inhibitors; Fluorescence; Src kinase; Density functional theory; Fourier transform infrared spectroscopy
• ISSN: 0003-7028; 1943-3530
• DOI: 10.1366/000370209788701125

The spectral properties of the SH2 and active site-directed sequences of the bivalent Src kinase inhibitor Ac-EELL(F5)Phe-(GABA)3-pYEEIE-amide (1) have been determined. Ac-pYEEIE-amide (2) and Ac-EELL(F5)Phe-amide (3), as well as the amino acids phosphotyrosine (pTyr) and pentafluorophenylalanine (F5)Phe, have been characterized by electronic absorption, fluorescence, and vibrational spectroscopy. Specific and unique marker bands that originate from the phosphate group of pTyr and the fluorinated aromatic ring of (F5)Phe have been identified, with the latter showing some solvent dependence. Peptide 2 was found to have excitation and emission wavelengths emanating from pTyr at 268 and 295 nm, respectively, whereas peptide 3 displayed excitation and emission peaks attributable to (F5)Phe at 274 and 315 nm, respectively. Fourier transform infrared (FT-IR) analysis of the amino acid pTyr identified distinct marker bands at approximately 930, 1090, and 1330 cm−1 that could be attributed to the phosphate group. These markers were also observed in the IR spectrum of peptide 2. Likewise, peptide 3 displayed a characteristic C–F stretching mode at 961 cm−1 due to the presence of (F5)Phe, including two C–F reporting ring modes at 1509 and 1527 cm−1. Identifying and monitoring spectroscopic changes in these marker bands may afford a means to observe the molecular interactions that occur when peptides 1–3 bind to the Src kinase.