Investigation of the phenylalanyl‐tRNA synthetase modification with γ‐(<i>p</i>‐Azidoanilide)‐ATP

Ankilova, V.N.; Knorre, D.G.; Kravchenko, Vladimir V.; Lavrik, Olga I.; Nevinsky, G.A.

doi:10.1016/0014-5793(75)80445-5

Cited by 20 publications

(5 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A potent photoaffinity marker, adenosine 5'-[y-(p-azidoanilido)]triphosphate has been synthesized by Budker et al [29] and used by Akhverdyan et al [30] to label tryptophanyl-tRNA synthetase. But in the case of E. coli phenylalanyl-tRNA synthetase, Ankilova et al [31] showed that the observed labelling was not specific. We thus turned to the acetone photosensitized crosslinking technique described by Sperling et al [32 -341.…”

Section: Photosensitized Cross-linking Of Atp To Phenylalanyl-trna Symentioning

confidence: 99%

Yeast Phenylalanyl‐tRNA Synthetase

Baltzinger

Fasiolo

Rémy

1979

European Journal of Biochemistry

View full text Add to dashboard Cite

The localization of the binding sites of the different ligands on the constitutive subunits of yeast phenylalanyl-tRNA synthetase was undertaken using a large variety of affinity and photoaffinity labelling techniques. The tRNAPh' was cross-linked to the enzyme by non-specific ultraviolet irradiation at 248 nm, specific irradiation in the wye base absorption band (315 nm), irradiation at 335 nm, in the absorption band of 4-thiouridine (s"U) residues introduced in the tRNA molecule, or by Schiffs base formation between periodate-oxidized tRNAPh' (tRNA;:') and the protein. ATP was specifically incorporated in its binding site upon photosensitized irradiation.The amino acid could be linked to the enzyme upon ultraviolet irradiation, either in the free state, engaged in the adenylate or bound to the tRNA.The tRNA, the ATP molecule and the amino acid linked to the tRNA were found to interact exclusively with the 0 subunit ( M , 63000). The phenylalanine residue, either free or joined to the adenylate, could be cross-linked with equal efficiency to either type of subunit, suggesting that the amino acid binding site is located in a contact area between the two subunits. The Schiffs base formation between tRNA:,he and the enzyme shows the existence of a lysyl group close to the binding site for the 3'-terminal adenosine of tRNA. This result was confirmed by the study of the inhibition of yeast phenylalanyl-tRNA synthetase with pyridoxal phosphate and the 2',3'-dialdehyde derivative of ATP, oATP.

show abstract

Section: Photosensitized Cross-linking Of Atp To Phenylalanyl-trna Symentioning

confidence: 99%

Yeast Phenylalanyl‐tRNA Synthetase

Baltzinger

Fasiolo

Rémy

1979

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…TPLN synthesis 15-17 is simple compared to that of many other nucleotide analogs, and the same conjugation strategy can be applied to all four nucleotides 10,11,14 . Although TPLFNs, in which the label is attached to the γ-phosphate, are rather poor substrates for DNA polymerase 10,11 , extension of the phosphate chain to increase the distance between the labeling moiety and the polymerase active site, and recover the native charge of a nucleotide triphosphate has led to significant improvements 10,11 .…”

Section: Resultsmentioning

confidence: 99%

Fluorogenic DNA sequencing in PDMS microreactors

et al. 2011

View full text Add to dashboard Cite

We have developed a multiplex sequencing-by-synthesis method combining terminal-phosphate labeled fluorogenic nucleotides (TPLFNs) and resealable microreactors. In the presence of phosphatase, the incorporation of a non-fluorescent TPLFN into a DNA primer by DNA polymerase results in a fluorophore. We immobilize DNA templates within polydimethylsiloxane (PDMS) microreactors, sequentially introduce one of the four identically labeled TPLFNs, seal the microreactors, allow template-directed TPLFN incorporation, and measure the signal from the fluorophores trapped in the microreactors. This workflow allows sequencing in a manner akin to pyrosequencing but without constant monitoring of each microreactor. With cycle times of <10 minutes, we demonstrate 30 base reads with ∼99% raw accuracy. “Fluorogenic pyrosequencing” combines benefits of pyrosequencing, such as rapid turn-around, native DNA generation, and single-color detection, with benefits of fluorescence-based approaches, such as highly sensitive detection and simple parallelization.

show abstract

“…used for affinity labeling [2,3]. Available methods for the preparation of these compounds involve multistage procedures providing low or moderate yields [5-81.…”

Section: ' P-o 'P-o' 8 Lo-mentioning

confidence: 99%