Conformational restraint is a critical determinant of unnatural nucleotide recognition by protein kinases

• Published in: Bioorganic & medicinal chemistry letters Vol. 12; no. 21; pp. 3223 - 3227
• Main Authors: Ulrich, Scott M, Sallee, Nathan A, Shokat, Kevan M
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 04.11.2002; Elsevier
• Subjects: Adenosine Triphosphate - analogs & derivatives; Adenosine Triphosphate - metabolism; Adenosine Triphosphate - pharmacology; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Humans; Hydrogen Bonding; Mitogen-Activated Protein Kinases - chemistry; Mitogen-Activated Protein Kinases - metabolism; Molecular Conformation; Mutation; Nucleotides - metabolism; p38 Mitogen-Activated Protein Kinases; Protein Kinases - genetics; Protein Kinases - metabolism; Ribavirin - analogs & derivatives; Structure-Activity Relationship; Substrate Specificity; Transferases; Tyrosine - metabolism; Imidazole derivatives; Phosphorylation; Enzyme; Aminoamide; Transferases; Triphosphates; In vitro; Biological activity; Signal transduction; Enzymatic activity; Protein kinase; Nucleotide; Chemical synthesis; Ribonucleotide
• ISSN: 0960-894X; 1464-3405
• DOI: 10.1016/S0960-894X(02)00616-9

This report describes the synthesis of N 4-(benzyl) AICAR triphosphate, a conformationally restrained analogue of N 4-(benzyl) ribavirin triphosphate. Both of these nucleotides were evaluated as phosphodonors for wild-type p38 MAP kinase and T106G p38 MAP kinase, a designed mutant with expanded nucleotide specificity. The conformationally restrained nucleotide, N 4-(benzyl) AICAR triphosphate, is orthogonal to (not accepted as a substrate by) wild-type p38 MAP kinase, in contrast to N 4-(benzyl) ribavirin triphosphate. Furthermore, N 4-(benzyl) AICAR triphosphate, is accepted as a substrate by T106G p38 MAP kinase, in contrast to N 4-(benzyl) ribavirin triphosphate. We hypothesize that the presence of an internal hydrogen bond in N 4-(benzyl) AICAR and its absence in N 4-(benzyl) ribavirin triphosphate is the main determinant for their differing structure–activity relationships. Graphic