Biotin-c10-AppCH2ppA is an effective new chemical proteomics probe for diadenosine polyphosphate binding proteins.
Here we report on the synthesis of a synthetic, stable biotin-c10-AppCH2ppA conjugate involving an unusual Cannizzaro reaction step. This conjugate is used to bind prospective Ap4A binding proteins from Escherichia coli bacterial cell lyzates. Following binding, identities of these proteins are then determined smoothly by a process of magnetic bio-panning and electrospray mass spectrometry. Protein hits appear to be a definitive set of stress protein related targets. While this hit list may not be exclusive, and may vary with the nature of sampling conditions and organism status, nevertheless hits do appear to correspond with bona fide Ap4A-binding proteins. Therefore these hits represent a sound basis on which to construct new hypotheses concerning the cellular importance of Ap4A to bacterial cells and the potential biological significance of Ap4A-protein binding interactions.
Imperial College Genetic Therapies Centre, Department of Chemistry, Flowers Building, Armstrong Road, Imperial College London, London SW7 2AZ,UK; Division of Organic Chemistry, Indian Institute of Chemical Technology, Hyderabad, India.
M Ameruddin
MA
Azhar
Imperial College Genetic Therapies Centre, Department of Chemistry, Flowers Building, Armstrong Road, Imperial College London, London SW7 2AZ,UK; Institute of Pharmaceutical Science, King's College London, Franklin-Wilkins Building, Waterloo Campus, 150 Stamford Street, London SE1 9NH,UK.
Michael
M
Wright
Division of Organic Chemistry, Indian Institute of Chemical Technology, Hyderabad, India.
Ahmed
A
Kamal
Institute of Biomedical Engineering, Armstrong Road, Imperial College London, London SW7 2AZ,UK.
Judith
J
Nagy
Imperial College Genetic Therapies Centre, Department of Chemistry, Flowers Building, Armstrong Road, Imperial College London, London SW7 2AZ,UK; Institute of Pharmaceutical Science, King's College London, Franklin-Wilkins Building, Waterloo Campus, 150 Stamford Street, London SE1 9NH,UK. Electronic address: a.miller07@btinternet.com.
Andrew D
AD
Miller
D000225:Adenine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D002352:Carrier Proteins / Q000032:analysis*; D015226:Dinucleoside Phosphates / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry / Q000166:cytology; D008968:Molecular Conformation; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry*; D040901:Proteomics
D002352:Carrier Proteins; D015226:Dinucleoside Phosphates; D015335:Molecular Probes; C020733:diadenosine tetraphosphate; D001710:Biotin; D000225:Adenine
Ap(4)A binding proteins; Chemical proteomics; Diadenosine tetraphosphate analogues; Dinucleoside polyphosphates; Escherichia coli