Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or...
Reexamination Certificate
2007-08-14
2007-08-14
Saucier, Sandra E. (Department: 1651)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
C435S015000
Reexamination Certificate
active
10888378
ABSTRACT:
High-Throughput Screening (HTS) of large compound libraries is the method of drug-lead discovery. It is now well accepted that for a functional assay, quality is more important than quantity. A biochemical NMR method originally proposed by Percival and Withers (Biochemistry, 1992, 31, 498–505) is extended to the screening of Ser/Thr kinases. The method requires the presence of a CF3(or CF) moiety on the substrate and utilizes19F NMR spectroscopy for the detection of the starting and enzymatically modified substrates. Experiments can be performed in real time or in an endpoint assay format using protein and substrate concentrations comparable to the ones used by other HTS techniques. Application of this technique to the phosphorylation of a substrate by the protein Ser/Thr kinase AKT1 is presented.
REFERENCES:
On-Line Medical Dictionary definition for “deconvolution”.
Reuveni et al., “Toward a PKB Inhibitor: Modification of a Selective PKA Inhibitor by Rational Design”, Biochemistry 41: 10304-10314 (2002).
Percival et al., “19F NMR Investigations of the Catalytic Mechanism of Phosphoglucomutase Using Fluorinated Substrates and Inhibitors”, Biochemisty 31: 505-512 (1992).
Ardini Elena
Dalvit Claudio
Flocco Maria Magdalena
Fogliatto Gianpaolo
Mongelli Nicola
Pfizer Italia S.r.l.
Saucier Sandra E.
Scully , Scott, Murphy & Presser, P.C.
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