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A Parallel Synthesis Scheme for Generating Libraries of DNA Polymerase Substrates and Inhibitors
Author(s) -
Strobel Heike,
Dugué Laurence,
Marlière Philippe,
Pochet Sylvie
Publication year - 2002
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/1439-7633(20021202)3:12<1251::aid-cbic1251>3.0.co;2-1
Subject(s) - chemistry , nucleoside triphosphate , stereochemistry , polymerase , dna polymerase , dna , primer (cosmetics) , dna polymerase i , nucleotide , combinatorial chemistry , imidazole , pyrimidine , biochemistry , polymerase chain reaction , organic chemistry , reverse transcriptase , gene
We report a combinatorial approach aimed at producing in a single step a large family of nucleoside triphosphate derivatives that could be tested for their ability to be substrates for DNA polymerases. We propose as a unique triphosphate building block a nucleotide with a hydrazine function anchored to an imidazole ring. Condensation between the 5′‐triphosphate derivative of 1‐(2‐deoxy‐ β ‐ D ‐ erythro ‐pentofuranosyl)‐imidazole‐4‐hydrazide (dY NH 2TP) and any aldehyde or ketone, followed by reduction of the intermediate hydrazones dXmTP, resulted in the corresponding hydrazides (dXnTP). Following this scheme, a series of aldehydes having various aromatic parts yielded a number of adducts dY NHR TP. Vent (exo − ) DNA polymerase is found to be able to catalyse the single incorporation of these bulky triphosphate derivatives. Subsequent extensions of the modified pairs with canonical triphosphates resulted mainly in abortive elongations at primer+2, except after the incorporation of dY NHben TP and, to a lesser extent, dY NHphe TP opposite C. These results illustrate the potential of this parallel synthetic scheme for generating new substrates or inhibitors of replication in a single step.