J 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

Štítky

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
Název: Vývoj metodologie s vysokým rozlišením pro NMR studie neuspořádaných proteinů s vysoce degenerovanými rezonančními frekvencemi (Akronym: HIGHRES)
Investor: Grantová agentura ČR, Vývoj metodologie s vysokým rozlišením pro NMR studie neuspořádaných proteinů s vysoce degenerovanými rezonančními frekvencemi
228461, interní kód MU
Název: Enhancing Access and Services to East European users Towards an efficient (Akronym: EAST-NMR)
Investor: Evropská unie, Enhancing Access and Services to East European users Towards an efficient, Kapacity