Enzyme Family Specific and Activity Based Screening of Chemical Libraries Using Enzyme Microarrays  

Oleh:

Funeriu, Daniel P. ; Eppinger, Jorg ; Denizot, Lucile ; Miyake, Masato ; Miyake, Jun

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Nature Biotechnology: The Science and Business of Biotechnology vol. 23 no. 5 (Mei 2005), page 622-627.
Topik: enzymes; enzyme; chemical; microarrays

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: NN9.3 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

Lihat Detail Induk

Isi artikel

The potential of protein microarrays in high throughput screening (HTS) still remains largely unfulfilled, essentially because of the difficulty of extracting meaningful, quantitative data from such experiments. In the particular case of enzyme microarrays, low molecular weight fluorescent affinity labels (FALs) can function as ideally suited activity probes of the microarrayed enzymes. FALs form covalent bonds with enzymes in an activity dependent manner and therefore can be used to characterize enzyme activity at each enzymes address, as predetermined by microarraying process. Relying on this principle, we introduce herein thematic enzyme microarrays (TEMA). In a kinetic setup we used TEMAs to determine the full set of kinetic constants and the reaction mechanism between the microarrayed enzymes (the theme of the microarray) and a family wide FAL. Based on this kinetic understanding, and HTS setup we established the practical and theoretical methodology for quantitative, multiplexed determination of the inhibition profile of compounds from a chemical library against each microarrayed enzyme. Finally, in a validation setup Ki (app) values and inhibitor profiles were confirmed and refined.

Opini Anda

Klik untuk menuliskan opini Anda tentang koleksi ini!

Kembali