Anti-sulfonylbenzoate antibodies as a tool for the detection of nucleotide-binding proteins for functional proteomics.
Proteins that bind ATP and GTP are important cellular components. We developed an immunological approach to selectively tag nucleotide-binding proteins based on the use of 5'-[4-(fluorosulfonyl)benzoyl]adenosine and 5'-[4-(fluorosulfonyl)benzoyl]guanosine affinity tags and an antibody against 4-(sulfonyl)benzoate. Detection follows affinity labeling, gel electrophoresis, and ester bond cleavage to expose the epitope. Trial analyses of labeled proteins from lymphoid cells identified multiple ATP-binding proteins, including chaperones, actin, kinases, an RNA splicing factor, a membrane ATPase, and ATP synthase.
Moore
Lisa L
LL
Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN 47907, USA.
Fulton
Ashley M
AM
Harrison
Marietta L
ML
Geahlen
Robert L
RL
D000255:Adenosine Triphosphate / Q000378:metabolism; D000345:Affinity Labels; D000911:Antibodies, Monoclonal; D001402:B-Lymphocytes; D001565:Benzoates / Q000276:immunology*; D002352:Carrier Proteins / Q000032:analysis* / Q000378:metabolism; D002457:Cell Extracts / Q000737:chemistry; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D008214:Lymphocytes / Q000737:chemistry; D009711:Nucleotides / Q000378:metabolism; D040901:Proteomics / Q000379:methods*
D000345:Affinity Labels; D000911:Antibodies, Monoclonal; D001565:Benzoates; D002352:Carrier Proteins; D002457:Cell Extracts; D009711:Nucleotides; C084636:4-(fluorosulfonyl)benzoic acid; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate