N-hydroxysuccinimide ester functionalized perfluorophenyl azides as novel photoactive heterobifunctional cross-linking reagents. The covalent immobilization of biomolecules to polymer surfaces.
AuthorsYan M, Cai SX, Wybourne MN, Keana JF
JournalBioconjug Chem
PubMed ID8031878
The synthesis of N-hydroxysuccinimide (NHS) functionalized perfluorophenyl azides (PFPAs) 2 and 3 is described together with a general method for the covalent modification of polymer surfaces using heterobifunctional, photoactivable cross-linking reagents 1 and 3. The NHS-active ester group becomes covalently attached to the polymer surface via an efficient CH bond ... More
Site-specific photomodification of single-stranded DNA targets by arylazide and perfluoroarylazide derivatives of oligonucleotides.
'Highly efficient site-specific photomodification of single-stranded DNA targets was achieved with oligonucleotide reagents bearing aromatic azido groups (R (R1 = p-azidotetrafluorobenzoyl, R2 = 2-nitro-5-azidobenzoyl, R3 = p-azidobenzoyl) at either the terminal phosphate or at the C5 position of deoxyuridine at the end or inside of the oligonucleotide chain. The extent ... More
Structure of photoreactive binary system of oligonucleotide conjugates assembled on the target nucleotide sequence.
AuthorsDobrikov MI, Bichenkova EV, Douglas KT, Gainutdinov TI, Vlassov VV
JournalJ Biomol Struct Dyn
PubMed ID10563571
'Recently we have developed an approach to superspecific photomodification of nucleic acids by binary systems of oligonucleotides conjugated to precursor groups capable of assembling into photoactivatable structure upon simultaneous binding of the conjugates to the target. We have investigated the solution structure of a model binary system 1:2:3, where 1 ... More
Application of an N-(4-azido-2,3,5,6-tetrafluorobenzoyl)tyrosine-substituted peptide as a heterobifunctional cross-linking agent in a study of protein O-glycosylation in yeast.
AuthorsDrake RR, Slama JT, Wall KA, Abramova M, D'Souza C, Elbein AD, Crocker PJ, Watt DS
JournalBioconjug Chem
PubMed ID1616952
'In order to investigate the O-mannosyltransferase involved in the initial O-mannosylation of glycoproteins in Saccharomyces cerevisiae, a photoactive hexapeptide, [125I]-N-(4-azido-2,3,5,6-tetrafluorobenzoyl)-3-iodo-Tyr-Asn-Pro-T hr-Ser-Val ([125I]azidoTyr-peptide), was synthesized by solid-phase techniques using a new photoactive cross-linking reagent, N-(4-azido-2,3,5,6-tetrafluorobenzoyl)tyrosine, and resin-bound Asn-Pro-Thr(tBu)-Ser(tBu)-Val. When this modified hexapeptide substrate was incubated with O-mannosyltransferase preparations, the hexapeptide was ... More
Functional organization of two large subunits of the fission yeast Schizosaccharomyces pombe RNA polymerase II. Location of the catalytic sites.
AuthorsWlassoff WA, Kimura M, Ishihama A
JournalJ Biol Chem
PubMed ID9988759
'The catalytically competent transcription complex of RNA polymerase II from the fission yeast Schizosaccharomyces pombe was affinity labeled with photoreactive nucleotide analogues incorporated at 3'' termini of nascent RNA chains. To locate the catalytic site for RNA polymerization, the labeled subunits were separated by SDS-polyacrylamide gel electrophoresis and subjected to ... More
Photoaffinity labelling of cyanomethaemoglobin with derivatives of tryptophan and 5-bromotryptophan.
AuthorsLi M, Lin Z, Johnson ME
JournalBiochem J
PubMed ID7755572
'Tryptophan and 5-bromotryptophan (5-BrTrp) are relatively potent inhibitors of sickle-haemoglobin polymerization. The binding sites of these compounds to normal and sickle haemoglobin (HBA and HBS) have been suggested, but not firmly established, through the use of spin-labelled derivatives and/or computer modeling. In the present study we approached the problem by ... More
Photoreactive insulin derivatives for the detection of the doubly labeled insulin receptor.
AuthorsKleinjung J, Fabry M
JournalPeptides
PubMed ID10793223
Two different photoinsulins, the radioactive N(epsilonB29)-(4-azidosalicyloyl) insulin and the novel biotinylated N(epsilonB29)-(4-azidotetrafluorobenzoyl-biocytinyl) insulin, were synthesized in order to study the binding stoichiometry of insulin and the insulin receptor in a direct approach. Both derivatives were cross-linked simultaneously to the (alphabeta)(2) receptor. Insulin-receptor conjugates were formed that carry a radioactive label ... More
A novel fiber-optic pH sensor incorporating carboxy SNAFL-2 and fluorescent wavelength-ratiometric detection.
AuthorsXu Z, Rollins A, Alcala R, Marchant RE
JournalJ Biomed Mater Res
PubMed ID9429091
A fiber-optic pH sensor was prepared using the self-referencing fluorescent pH probe carboxy seminaphtho-fluorescein (SNAFL-2). The ratio of the emission from the base form of this dye and the emission near the isoemissive point is insensitive to variations in the excitation intensity and photobleaching. The carboxy SNAFL-2 and a photoaffinity ... More
Oligonucleotide derivatives bearing reactive and stabilizing groups attached to C5 of deoxyuridine.
Oligonucleotides bearing an aliphatic amino group at the C5-position of deoxyuridine (ULNH2TCCCA, TULNH2CCCA, ULNH2CCACTT, where L = -CH2-, -CH2OCH2CH2- or -CH2NHCOCH2CH2-) have been synthesized. The photoactive (p-azidotetrafluorobenzamido, 2-nitro-5-azidobenzamido, or p-azidobenzamido), alkylating [4-[N-(2-chloroethyl)-N-methylamino]benzyl], or intercalating [N-(2-hydroxyethyl)phenazinium] groups were attached to the amino linker of oligonucleotides. The Tm values were determined for ... More
5-[3-(E)-(4-azido-2,3,5,6-tetrafluorobenzamido)propenyl-1]-2'-deoxy- uridine-5'-triphosphate substitutes for thymidine-5'-triphosphate in the polymerase chain reaction.
AuthorsGodovikova TS, Kolpashchikov DM, Orlova TN, Richter VA, Ivanova TM, Grochovsky SL, Nasedkina TV, Victorova LS, Poletaev AI
JournalBioconjug Chem
PubMed ID10346887
The DNA targets may be labeled and simultaneously amplified in the polymerase chain reaction (PCR) using a pair of respective primers after elongation with nucleoside-5'-triphosphates carrying photoreactive groups. The amplified DNA may be subsequently photoactivated by irradiation above 300 nm, resulting in photo-cross-linking of the strands. For this goal 5-[3-(E)-(4-azido-2,3,5,6-tetrafluorobenzamido)propenyl-1]-, ... More
Survey of four different photoreactive moieties for DNA photoaffinity labeling of yeast RNA polymerase III transcription complexes.
AuthorsTate JJ, Persinger J, Bartholomew B
JournalNucleic Acids Res
PubMed ID9490787
In order to optimize the detection of protein-DNA contacts by DNA photoaffinity labeling, we attached four different photoreactive groups to DNA and examined their ability to crosslink yeast RNA polymerase III (Pol III) transcription complexes. Photoreactive nucleotides containing an aryl azide (AB-dUMP), benzophenone (BP-dUMP), perfluorinated aryl azide (FAB-dUMP) or diazirine ... More
Binary system for selective photoaffinity labeling of base excision repair DNA polymerases.
AuthorsLavrik OI, Kolpashchikov DM, Prasad R, Sobol RW, Wilson SH
JournalNucleic Acids Res
PubMed ID12136121
A system of photoaffinity reagents for selective labeling of DNA polymerases in extracts has been examined. To create the photoreactive DNA probe in situ, DNA substrates containing a synthetic abasic site are incubated in mouse embryonic fibroblast (MEF) cellular extract in the presence of base-substituted arylazido derivatives of dNTPs. This ... More
Structural studies by high-field NMR spectroscopy of a binary-addressed complementary oligonucleotide system juxtaposing pyrene and perfluoro-azide units.
AuthorsBichenkova EV, Marks DS, Lokhov SG, Dobrikov MI, Vlassov VV, Douglas KT
JournalJ Biomol Struct Dyn
PubMed ID9399157
Recently, a new approach has been proposed to improve the site-specificity and efficiency of the modification of nucleic acid target sequences, the binary system of complementary-addressing nucleic acid sequences. The binary system comprises two oligonucleotides, one modified with a photosensitizing group and the other with a photoreactive group. The sites ... More
Labeling of Torpedo californica nicotinic acetylcholine receptor subunits by cobratoxin derivatives with photoactivatable groups of different chemical nature at Lys23.
AuthorsUtkin YN, Krivoshein AV, Davydov VL, Kasheverov IE, Franke P, Maslennikov IV, Arseniev AS, Hucho F, Tsetlin VI
JournalEur J Biochem
PubMed ID9578481
Different photoactivatable derivatives of toxin 3 (CTX) Naja naja siamensis were obtained after CTX reaction with N-hydroxysuccinimide esters of p-azidobenzoic, p-azidotetraflourobenzoic, p-benzoylbenzoic and p-[3-(trifluoromethyl)-3H-diazirin-3-yl]benzoic acids. The ion-exchange HPLC profiles for the reaction products were very similar in four cases, with one predominant peak corresponding to the derivative containing the label ... More