[New reagents for affinity modification of biopolymers. Photoaffinity modification of Tte-DNA polymerase].
Arylazides N-(4-azido-2,5-difluoro-3-chloropyridinyl-6)-beta-alanine (Ia) and N-(4-azido-2,5-difluoro-3-chloropyridinyl-6)-glycine (Ib) were synthesized and covalently attached to 5-(3-aminopropenyl-1)-dUTP through the amino group to give 5'-triphosphate (IIa) and 5'-triphosphate (IIb). The resulting azides were subjected to photolysis in aqueous solution. The spectral and photochemical characteristics of azides (I) and (II) imply that their use for the modification of biopolymers holds promise. Compounds (IIa, b) effectively substituted dTTP in DNA polymerization catalyzed by thermostable DNA polymerase from Thermus thermophilus B-35 (Tte DNA polymerase). Photoaffinity modification of Tte DNA polymerase was carried out by dTTP analogues (IIa, b) and by earlier obtained 5-[N-(5-azido-2-nitrobenzoyl)-trans-3-aminopropenyl-1]deoxyuridine 5'-triphosphate (III) and 5-[N-(4-azido-2,3,5,6-tetrafluorobenzyol)-trans-3- aminopropenyl-1]deoxyuridine 5'-triphosphate (IV) using two variants of labeling. All four dTTP analogues were shown to modify Tte DNA polymerase.
Kolpashchikov
D M
DM
Novosibirsk Institute of Bioorganic Chemistry, Siberian Division, Russian Academy of Sciences, Russia.
Zakharenko
A L
AL
Dezhurov
S V
SV
Rechkunova
N I
NI
Khodyreva
S N
SN
Degtiarev
S Kh
SKh
Litvak
V V
VV
Lavrik
O I
OI
D001386:Azides / Q000138:chemical synthesis*; D001483:Base Sequence; D001704:Biopolymers / Q000737:chemistry*; D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry*; D003856:Deoxyuracil Nucleotides / Q000737:chemistry; D005998:Glycine / Q000031:analogs & derivatives / Q000138:chemical synthesis*; D019920:Photoaffinity Labels; D013056:Spectrophotometry, Ultraviolet; D016963:Thermus thermophilus / Q000201:enzymology; D013942:Thymine Nucleotides / Q000737:chemistry; D015091:beta-Alanine / Q000031:analogs & derivatives / Q000138:chemical synthesis*
D001386:Azides; D001704:Biopolymers; D017931:DNA Primers; D003856:Deoxyuracil Nucleotides; D019920:Photoaffinity Labels; D013942:Thymine Nucleotides; C027078:deoxyuridine triphosphate; D015091:beta-Alanine; D004259:DNA-Directed DNA Polymerase; C024157:thymidine 5'-triphosphate; D005998:Glycine