2010
Strategy for complete NMR assignment of disordered proteins with highly repetitive sequences based on resolution-enhanced 5D experiments
MOTÁČKOVÁ, Veronika; Jiří NOVÁČEK; Anna ZAWADZKA-KAZIMIERCZUK; Krzysztof KAZIMIERCZUK; Lukáš ŽÍDEK et. al.Základní údaje
Originální název
Strategy for complete NMR assignment of disordered proteins with highly repetitive sequences based on resolution-enhanced 5D experiments
Název česky
Strategie pro kompletní NMR přiřazení nestrukturovaných proteinů s vysokým počtem repeticí v sekvenci
Autoři
MOTÁČKOVÁ, Veronika (203 Česká republika, garant, domácí); Jiří NOVÁČEK (203 Česká republika, domácí); Anna ZAWADZKA-KAZIMIERCZUK (616 Polsko); Krzysztof KAZIMIERCZUK (616 Polsko); Lukáš ŽÍDEK (203 Česká republika, domácí); Hana ŠANDEROVÁ (203 Česká republika); Libor KRÁSNÝ (203 Česká republika); Wiktor KOZMINSKI (616 Polsko) a Vladimír SKLENÁŘ (203 Česká republika, domácí)
Vydání
Journal of Biomolecular NMR, Netherlands, Springer Netherlands, 2010, 0925-2738
Další údaje
Jazyk
angličtina
Typ výsledku
Článek v odborném periodiku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Nizozemské království
Utajení
není předmětem státního či obchodního tajemství
Odkazy
Impakt faktor
Impact factor: 3.047
Kód RIV
RIV/00216224:14310/10:00045112
Organizační jednotka
Přírodovědecká fakulta
UT WoS
000283373600006
Klíčová slova česky
Unfolded proteins; Repetitive sequence; Multi-dimensional NMR; Non-uniform sampling; Assignment
Klíčová slova anglicky
Unfolded proteins; Repetitive sequence; Multi-dimensional NMR; Non-uniform sampling; Assignment
Příznaky
Mezinárodní význam, Recenzováno
Změněno: 2. 2. 2012 17:33, Mgr. Veronika Papoušková, Ph.D.
V originále
A strategy for complete backbone and side-chain resonance assignment of disordered proteins with highly repetitive sequence is presented. The protocol is based on three resolution-enhanced NMR experiments: 5D HN(CA)CONH provides sequential connectivity, 5D HabCabCONH is utilized to identify amino acid types, and 5D HC(CC-TOCSY)CONH is used to assign the side-chain resonances. The improved resolution was achieved by a combination of high dimensionality and long evolution times, allowed by non-uniform sampling in the indirect dimensions. Random distribution of the data points and Sparse Multidimensional Fourier Transform processing were used. Successful application of the assignment procedure to a particularly difficult protein, delta subunit of RNA polymerase from Bacillus subtilis, is shown to prove the efficiency of the strategy. The studied protein contains a disordered C-terminal region of 81 amino acids with a highly repetitive sequence. While the conventional assignment methods completely failed due to a very small differences in chemical shifts, the presented strategy provided a complete backbone and side-chain assignment.
Česky
A strategy for complete backbone and side-chain resonance assignment of disordered proteins with highly repetitive sequence is presented. The protocol is based on three resolution-enhanced NMR experiments: 5D HN(CA)CONH provides sequential connectivity, 5D HabCabCONH is utilized to identify amino acid types, and 5D HC(CC-TOCSY)CONH is used to assign the side-chain resonances. The improved resolution was achieved by a combination of high dimensionality and long evolution times, allowed by non-uniform sampling in the indirect dimensions. Random distribution of the data points and Sparse Multidimensional Fourier Transform processing were used. Successful application of the assignment procedure to a particularly difficult protein, delta subunit of RNA polymerase from Bacillus subtilis, is shown to prove the efficiency of the strategy. The studied protein contains a disordered C-terminal region of 81 amino acids with a highly repetitive sequence. While the conventional assignment methods completely failed due to a very small differences in chemical shifts, the presented strategy provided a complete backbone and side-chain assignment.
Návaznosti
| GA204/09/0583, projekt VaV |
| ||
| GD301/09/H004, projekt VaV |
| ||
| LC06030, projekt VaV |
| ||
| MSM0021622413, záměr |
|