TY - JOUR
T1 - Dissection of bacteriophage λ site-specific recombination using synthetic peptide
AU - Cassell, Geoffrey
AU - Klemm, Martha
AU - Pinilla, Clemencia
AU - Segall, Anca
PY - 2000/6/23
Y1 - 2000/6/23
N2 - A wide variety of tools have been used to dissect biochemical pathways, inhibitors being chief among them. Combinatorial approaches have made the search for inhibitors much more efficient. We have applied such an approach to identify hexapeptides which inhibit different steps in a site-specific recombination reaction mediated by the bacteriophage λ integrase protein. Integrase's mechanism is still incompletely understood, in large part because several pathway intermediates remain hard to isolate. Integrase-catalyzed recombination is very efficient, but if blocked, it is highly reversible to substrates; this combination makes some intermediates exceedingly transient. We have used synthetic peptide combinatorial libraries to screen for hexapeptides that affect the recombination pathway at different stages, and have identified two families of peptides: One probably blocks DNA cleavage, the other may stabilize the Holliday junction intermediates. These peptides do not resemble parts of integrase or any of the other helper functions in the pathway. The deconvolution of hexapeptide libraries based both on inhibition of an enzymatic reaction as well as on accumulation of reaction intermediates is a novel approach to finding useful tools for dissecting a biochemical pathway. (C) 2000 Academic Press.
AB - A wide variety of tools have been used to dissect biochemical pathways, inhibitors being chief among them. Combinatorial approaches have made the search for inhibitors much more efficient. We have applied such an approach to identify hexapeptides which inhibit different steps in a site-specific recombination reaction mediated by the bacteriophage λ integrase protein. Integrase's mechanism is still incompletely understood, in large part because several pathway intermediates remain hard to isolate. Integrase-catalyzed recombination is very efficient, but if blocked, it is highly reversible to substrates; this combination makes some intermediates exceedingly transient. We have used synthetic peptide combinatorial libraries to screen for hexapeptides that affect the recombination pathway at different stages, and have identified two families of peptides: One probably blocks DNA cleavage, the other may stabilize the Holliday junction intermediates. These peptides do not resemble parts of integrase or any of the other helper functions in the pathway. The deconvolution of hexapeptide libraries based both on inhibition of an enzymatic reaction as well as on accumulation of reaction intermediates is a novel approach to finding useful tools for dissecting a biochemical pathway. (C) 2000 Academic Press.
KW - Bacteriophage λ integrase
KW - Combinatorial chemistry
KW - Holliday junction
KW - Inhibitory peptides
KW - Tyrosine recombinase
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U2 - 10.1006/jmbi.2000.3828
DO - 10.1006/jmbi.2000.3828
M3 - Article
C2 - 10873445
AN - SCOPUS:0034705344
SN - 0022-2836
VL - 299
SP - 1193
EP - 1202
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 5
ER -