Fragmentation study on butanolides with tandem mass spectrometry and its application for the screening of ScbR-captured quorum sensing molecules in Streptomyces coelicolor A3(2).
Streptomyces coelicolor has a quorum sensing (QS) system triggered by small diffusible signaling molecules, i.e. butanolides (or gamma-butyrolactones) and their cognate DNA-binding receptors. Using the DNA-binding receptors as an affinity capture matrix, the butanolides can be easily enriched and identified. For the identification and screening of the butanolides, the diagnostic peak lists generated by the tandem mass spectrometric (MS/MS) fragmentation analysis of chemically synthetic butanolides were used. In the case of using ScbR as the capture matrix, SCB1, a previously well-known butanolide, and Acl-1 (or SCB3)-type butanolides having one more carbon in the acyl chain than SCB1, were detected. This is the first report directly demonstrating that Acl-1 is able to bind to ScbR in S. coelicolor. Our proposed method using both diagnostic peak lists of butanolide and the purified receptor protein as an affinity capture tool can be applied to rapidly screen QS molecules in vitro.
Joo
Hwang-Soo
HS
School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Republic of Korea.
Yang
Yung-Hun
YH
Lee
Chang-Soo
CS
Kim
June-Hyung
JH
Kim
Byung-Gee
BG
D015107:4-Butyrolactone / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D053038:Quorum Sensing; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D048372:Streptomyces coelicolor / Q000737:chemistry* / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C446304:ScbR protein, Streptomyces coelicolor; D015107:4-Butyrolactone