TY - JOUR
T1 - High-performance liquid chromatography analysis of N-acyl homoserine lactone hydrolysis by paraoxonases.
AU - Teiber, John F.
AU - Draganov, Dragomir I.
PY - 2011
Y1 - 2011
N2 - Mammalian paraoxonases (PONs) are a unique, highly conserved family of calcium-dependent esterases consisting of PON1, PON2, and PON3. The PONs can hydrolyze the lactone ring of a range of N-acyl-L: -homoserine lactone (AHL) quorum sensing signaling molecules, rendering them inactive. This chapter describes a method that utilizes high-performance liquid chromatography analysis with UV detection for determining the rate of AHL hydrolysis in cell lysates, tissue homogenates, serum, and with purified proteins. Also described are the techniques used to prepare cell culture lysates and tissue homogenates for analysis and the use of class-specific enzyme inhibitors to determine the contribution of PONs to AHL hydrolysis in the samples.
AB - Mammalian paraoxonases (PONs) are a unique, highly conserved family of calcium-dependent esterases consisting of PON1, PON2, and PON3. The PONs can hydrolyze the lactone ring of a range of N-acyl-L: -homoserine lactone (AHL) quorum sensing signaling molecules, rendering them inactive. This chapter describes a method that utilizes high-performance liquid chromatography analysis with UV detection for determining the rate of AHL hydrolysis in cell lysates, tissue homogenates, serum, and with purified proteins. Also described are the techniques used to prepare cell culture lysates and tissue homogenates for analysis and the use of class-specific enzyme inhibitors to determine the contribution of PONs to AHL hydrolysis in the samples.
UR - http://www.scopus.com/inward/record.url?scp=79952197625&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79952197625&partnerID=8YFLogxK
U2 - 10.1007/978-1-60761-971-0_21
DO - 10.1007/978-1-60761-971-0_21
M3 - Article
C2 - 21031320
AN - SCOPUS:79952197625
SN - 1064-3745
VL - 692
SP - 291
EP - 298
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
ER -