TY - JOUR
T1 - Screening of combinatorial libraries for substrate preference by mass spectrometry
AU - Stevens, Stanley M.
AU - Prokai-Tatrai, Katalin
AU - Prokai, Laszlo
PY - 2005/1/15
Y1 - 2005/1/15
N2 - We present a rapid screening method for monitoring enzyme specificity using both combinatorial chemistry and mass spectrometry where, as an example, the substrate specificity of peptidylglycine α-amidating enzyme was determined and compared against a conventional quantitative technique. Whereas alternative methods for library screening are generally limited to certain enzymes and can present difficulties in the synthesis or derivatization of potential substrates, the approach we call chirality-based isotope labeling for a library of substrates (CHILLS) does not fall short to such limitations, since we exploit the inherent stereospecificity of enzymes to determine preferred substrates. Additionally, the CHILLS method generates accurate results, as compared to typical screening procedures that require tedious method development, because the synthesized library contains a structurally similar internal standard for each individual library component in order to quantitate the progress of enzymatic reactions.
AB - We present a rapid screening method for monitoring enzyme specificity using both combinatorial chemistry and mass spectrometry where, as an example, the substrate specificity of peptidylglycine α-amidating enzyme was determined and compared against a conventional quantitative technique. Whereas alternative methods for library screening are generally limited to certain enzymes and can present difficulties in the synthesis or derivatization of potential substrates, the approach we call chirality-based isotope labeling for a library of substrates (CHILLS) does not fall short to such limitations, since we exploit the inherent stereospecificity of enzymes to determine preferred substrates. Additionally, the CHILLS method generates accurate results, as compared to typical screening procedures that require tedious method development, because the synthesized library contains a structurally similar internal standard for each individual library component in order to quantitate the progress of enzymatic reactions.
UR - http://www.scopus.com/inward/record.url?scp=12244280414&partnerID=8YFLogxK
U2 - 10.1021/ac0489925
DO - 10.1021/ac0489925
M3 - Article
C2 - 15649074
AN - SCOPUS:12244280414
SN - 0003-2700
VL - 77
SP - 698
EP - 701
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 2
ER -