title: A new functional, chemical proteomics technology to identify purine nucleotide binding sites in complex proteomes.
abstract: Adenine nucleotides are small, abundant molecules that bind numerous proteins involved in pivotal cellular processes. These nucleotides are co-factors or substrates for enzymes, regulators of protein function, or structural binding motifs. The identification of nucleotide-binding sites on a proteome-wide scale is tempting in view of the high number of nucleotide-binding proteins, their large in vivo concentration differences, and the various functions they exert. Here, we report on a functional, chemical, gel-free proteomics technology that allows the identification of protein adenine nucleotide-binding site(s) in cell lysates. Our technology uses a synthetic ATP analogue, 5'-p-fluorosulfonylbenzoyladenosine (FSBA), as an affinity/activity-based probe for nucleotide-binding sites. When applied on a cellular level, 185 different FSBA-labeled sites in a human Jurkat cell lysate were identified. Functional and structural aspects of the use of FSBA on a proteome-wide scale are discussed.
authors.affiliation: Department of Medical Protein Research and Biochemistry, Flanders Interuniversity Institute for Biotechnology and Faculty of Medicine and Health Sciences, Ghent University, A. Baertsoenkaai 3, B-9000 Ghent, Belgium.
authors.forename: Xavier
authors.initials: X
authors.lastname: Hanoulle
authors.forename: Jozef
authors.initials: J
authors.lastname: Van Damme
authors.forename: An
authors.initials: A
authors.lastname: Staes
authors.forename: Lennart
authors.initials: L
authors.lastname: Martens
authors.forename: Marc
authors.initials: M
authors.lastname: Goethals
authors.forename: Joël
authors.initials: J
authors.lastname: Vandekerckhove
authors.forename: Kris
authors.initials: K
authors.lastname: Gevaert
mesh_terms: D000225:Adenine / Q000378:metabolism*; D000241:Adenosine / Q000031:analogs & derivatives; D001665:Binding Sites / Q000235:genetics; D002457:Cell Extracts / Q000737:chemistry; D002853:Chromatography, Liquid; D006801:Humans; D019169:Jurkat Cells; D008958:Models, Molecular; D011506:Proteins / Q000235:genetics* / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D002457:Cell Extracts; D011506:Proteins; D020543:Proteome; C010959:5'-(4-fluorosulfonylbenzoyl)adenosine; D000225:Adenine; D000241:Adenosine