title: Design and synthesis of ATP-based nucleotide analogues and profiling of nucleotide-binding proteins.
abstract: Two nucleotide-based probes were designed and synthesized in order to enrich samples for specific classes of proteins by affinity-based protein profiling. We focused on the profiling of adenine nucleotide-binding proteins. Two properties were considered in the design of the probes: the bait needs to bind adenine nucleotide-binding proteins with high affinity and carry a second functional group suitable and easily accessible for coupling to a chromatography resin. For this purpose, we synthesized p-biotinyl amidobenzoic acid-ATP (p-BABA-ATP) and p-biotinyl aminomethylbenzoic acid-ATP (p-BAMBA-ATP). p-BABA-ATP and p-BAMBA-ATP both bind to ATP-binding cassette (ABC) proteins with at least 10-fold higher affinity than ATP. Several ABC transporters could be enriched using p-BABA-ATP or p-BAMBA-ATP.
authors.affiliation: Department of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute, Netherlands Proteomics Centre, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
authors.forename: Justina C
authors.initials: JC
authors.lastname: Wolters
authors.forename: Gerard
authors.initials: G
authors.lastname: Roelfes
authors.forename: Bert
authors.initials: B
authors.lastname: Poolman
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000227:Adenine Nucleotides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry*; D001105:Archaea / Q000378:metabolism*; D019843:Archaeal Proteins / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D013294:Lactococcus lactis / Q000378:metabolism*; D017015:Methanococcales / Q000378:metabolism; D008958:Models, Molecular; D011485:Protein Binding; D048229:Sulfolobus solfataricus / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; D000227:Adenine Nucleotides; D019843:Archaeal Proteins; D001426:Bacterial Proteins; D000255:Adenosine Triphosphate