An approach for high-throughput structure determination of proteins by NMR spectroscopy

Autor:	A, Medek, E T, Olejniczak, R P, Meadows, S W, Fesik
Rok vydání:	2001
Předmět:	Models Molecular Carbon Isotopes Proto-Oncogene Proteins c-bcl-2 bcl-X Protein Humans Proteins Nuclear Magnetic Resonance Biomolecular Sensitivity and Specificity Algorithms Protein Structure Tertiary
Zdroj:	Journal of biomolecular NMR. 18(3)
ISSN:	0925-2738
Popis:	An approach is described for rapidly determining protein structures by NMR that utilizes proteins containing 13C-methyl labeled Val, Leu, and Ile (delta1) and protonated Phe and Tyr in a deuterated background. Using this strategy, the key NOEs that define the hydrophobic core and overall fold of the protein are easily obtained. NMR data are acquired using cryogenic probe technology which markedly reduces the spectrometer time needed for data acquisition. The approach is demonstrated by determining the overall fold of the antiapoptotic protein, Bcl-xL, from data collected in only 4 days. Refinement of the Bcl-xL structure to a backbone rmsd of 0.95 A was accomplished with data collected in an additional 3 days. A distance analysis of 180 different proteins and structure calculations using simulated data suggests that our method will allow the global folds of a wide variety of proteins to be determined.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=pmid________::18ac96836151267b32a39342d94a44dc https://pubmed.ncbi.nlm.nih.gov/11142513 Zobrazit plný text záznamu Full text from SpringerLink