2013
NMR Determines Transient Structure and Dynamics in the Disordered C-Terminal Domain of WASp Interacting Protein
HABA, Noam Y, Renana GROSS, Jiří NOVÁČEK, Hadassa SHAKED, Lukáš ŽÍDEK et. al.Základní údaje
Originální název
NMR Determines Transient Structure and Dynamics in the Disordered C-Terminal Domain of WASp Interacting Protein
Autoři
HABA, Noam Y (376 Izrael), Renana GROSS (376 Izrael), Jiří NOVÁČEK (203 Česká republika, domácí), Hadassa SHAKED (376 Izrael), Lukáš ŽÍDEK (203 Česká republika, garant, domácí), Mira BARDA-SAAD (376 Izrael) a Jordan H CHILL (376 Izrael)
Vydání
Biophysical Journal, CAMBRIDGE, CELL PRESS, 2013, 0006-3495
Další údaje
Jazyk
angličtina
Typ výsledku
Článek v odborném periodiku
Obor
10600 1.6 Biological sciences
Stát vydavatele
Spojené státy
Utajení
není předmětem státního či obchodního tajemství
Odkazy
Impakt faktor
Impact factor: 3.832
Kód RIV
RIV/00216224:14740/13:00066376
Organizační jednotka
Středoevropský technologický institut
UT WoS
000321941700021
Klíčová slova anglicky
WISKOTT-ALDRICH-SYNDROME; NUCLEAR-MAGNETIC-RESONANCE; SEQUENCE-SPECIFIC ASSIGNMENT; PROLINE-RICH REGIONS; 8 M UREA; CHEMICAL-SHIFTS; ALPHA-SYNUCLEIN; N-WASP; ACTIN POLYMERIZATION; POLYPROLINE-II
Příznaky
Mezinárodní význam, Recenzováno
Změněno: 24. 10. 2013 09:18, prof. Mgr. Lukáš Žídek, Ph.D.
Anotace
V originále
WASp-interacting protein (WIP) is a 503-residue proline-rich polypeptide expressed in human T cells. The WIP C-terminal domain binds to Wiskott-Aldrich syndrome protein (WASp) and regulates its activation and degradation, and the WIP-WASp interaction has been shown to be critical for actin polymerization and implicated in the onset of WAS and X-linked thrombocytopenia. WIP is predicted to be an intrinsically disordered protein, a class of polypeptides that are of great interest because they violate the traditional structure-function paradigm. In this first (to our knowledge) study of WIP in its unbound state, we used NMR to investigate the biophysical behavior of WIPC, a C-terminal domain fragment of WIP that includes residues 407-503 and contains the WASp-binding site. In light of the poor spectral dispersion exhibited by WIPC and the high occurrence (25%) of proline residues, we employed 5D-(NMRC)-C-13-detected NMR experiments with nonuniform sampling to accomplish full resonance assignment. Secondary chemical-shift analysis, N-15 relaxation rates, and protection from solvent exchange all concurred in detecting transient structure located in motifs that span the WASp-binding site. Residues 446-456 exhibited a propensity for helical conformation, and an extended conformation followed by a short, capped helix was observed for residues 468-478. The C-13-detected approach allows chemical-shift assignment in the WIPC polyproline stretches and thus sheds light on their conformation and dynamics. The effects of temperature on chemical shifts referenced to a denatured sample of the polypeptide demonstrate that heating reduces the structural character of WIPC. Thus, we conclude that the disordered WIPC fragment is comprised of regions with latent structure connected by flexible loops, an architecture with implications for binding affinity and function.
Návaznosti
GAP206/11/0758, projekt VaV |
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228461, interní kód MU |
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