Immobilization of a novel antibacterial agent on solid phase and subsequent isolation of EF-Tu.
Screening of our compound collection identified PNU-92560, a 2-[1,3,4]thiadiazolo[3,2-a]pyrimidine-6-carboxamide, as a novel antibacterial agent. Extensive analogue development identified that the 2-position of the thiadiazole could be functionalized with a linker that would allow the compound to be attached to a solid support. The extreme insolubility of the analogues prevented the mechanism of action for these compounds to be determined utilizing traditional methodology. The solid-supported compounds were utilized as affinity columns to identify elongation factor Tu (EF-Tu) as a putative target for this class of compounds. The activity of the compounds in a metabolic labeling experiments and in translation assay supports the identity of the target for these compounds to be EF-Tu.
Deibel
Martin R
MR
Pharmacia, Kalamazoo, Michigan 49001-0199, USA. mrdeibel@proteos.net
Bodnar
Alice L
AL
Yem
Anthony W
AW
Wolfe
Cindy L
CL
Heckaman
Charlotte L
CL
Bohanon
Michael J
MJ
Mathews
W Rodney
WR
Sweeney
Michael T
MT
Zurenko
Gary E
GE
Marotti
Keith R
KR
Boyle
Timothy P
TP
Thorarensen
Atli
A
D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D016503:Drug Delivery Systems; D010444:Peptide Elongation Factor Tu / Q000302:isolation & purification* / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D011743:Pyrimidines / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013211:Staphylococcus aureus / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D013844:Thiazoles / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology
C493147:2-(1,3,4)thiadiazolo(3,2-a)pyrimidine-6-carboxamide; D000900:Anti-Bacterial Agents; D011743:Pyrimidines; D013844:Thiazoles; D010444:Peptide Elongation Factor Tu