title: Synthesis of two fluorescent GTPγS molecules and their biological relevance.
abstract: Fluorescent GTP analogues are utilized for an assortment of nucleic acid and protein characterization studies. Non-hydrolysable analogues such as GTPγS offer the advantage of keeping proteins in a GTP-bound conformation due to their resistance to hydrolysis into GDP. Two novel fluorescent GTPγS molecules were developed by linking fluorescein and tetramethylrhodamine to the γ-thiophosphate of GTPγS. Chemical and biological analysis of these two compounds revealed their successful synthesis and ability to bind to the nucleotide-binding site of tubulin. These two new fluorescent non-hydrolysable nucleotides offer new possibilities for biophysical and biochemical characterization of GTP-binding proteins.
authors.affiliation: a Department of Internal Medicine and UC Davis Comprehensive Cancer Center , University of California , Davis , CA , USA.
authors.forename: Denise J
authors.initials: DJ
authors.lastname: Trans
authors.affiliation: b Screening Technologies Branch, Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis, National Cancer Institute, Frederick National Laboratory for Cancer Research , National Institutes of Health , Frederick , MD , USA.
authors.forename: Ruoli
authors.initials: R
authors.lastname: Bai
authors.affiliation: c Nuclear Magnetic Resonance Facility , University of California , Davis , CA , USA.
authors.forename: J Bennet
authors.initials: JB
authors.lastname: Addison
authors.affiliation: d Department of Biochemistry and Molecular Medicine , University of California , Davis , CA , USA.
authors.forename: Ruiwu
authors.initials: R
authors.lastname: Liu
authors.affiliation: b Screening Technologies Branch, Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis, National Cancer Institute, Frederick National Laboratory for Cancer Research , National Institutes of Health , Frederick , MD , USA.
authors.forename: Ernest
authors.initials: E
authors.lastname: Hamel
authors.affiliation: e Department of Radiation Oncology , University of California , Davis , CA , USA.|f Lawrence Livermore National Laboratory , Livermore , CA , USA.
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Coleman
authors.affiliation: a Department of Internal Medicine and UC Davis Comprehensive Cancer Center , University of California , Davis , CA , USA.
authors.forename: Paul T
authors.initials: PT
authors.lastname: Henderson
mesh_terms: D060326:Chemistry Techniques, Synthetic; D004735:Energy Transfer; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000138:chemical synthesis* / Q000378:metabolism; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D014404:Tubulin / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D014404:Tubulin; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D019204:GTP-Binding Proteins
keywords: FRET; NMR and tubulin polymerization; Nucleoside and nucleotide analogs; fluorescent nucleotide analogs