NMR Determines Transient Structure and Dynamics in the
Disordered C-Terminal Domain of WASp Interacting Protein

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.

Basic information

Original name

NMR Determines Transient Structure and Dynamics in the Disordered C-Terminal Domain of WASp Interacting Protein

Authors

HABA, Noam Y (376 Israel), Renana GROSS (376 Israel), Jiří NOVÁČEK (203 Czech Republic, belonging to the institution), Hadassa SHAKED (376 Israel), Lukáš ŽÍDEK (203 Czech Republic, guarantor, belonging to the institution), Mira BARDA-SAAD (376 Israel) and Jordan H CHILL (376 Israel)

Edition

Biophysical Journal, CAMBRIDGE, CELL PRESS, 2013, 0006-3495

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 3.832

RIV identification code

RIV/00216224:14740/13:00066376

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1016/j.bpj.2013.05.046

UT WoS

000321941700021

Keywords in English

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

Abstract

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.

Links

GAP206/11/0758, research and development project

Name: Vývoj metodologie s vysokým rozlišením pro NMR studie neuspořádaných proteinů s vysoce degenerovanými rezonančními frekvencemi (Acronym: HIGHRES)

Investor: Czech Science Foundation

228461, interní kód MU

Name: Enhancing Access and Services to East European users Towards an efficient (Acronym: EAST-NMR)

Investor: European Union, Enhancing Access and Services to East European users Towards an efficient, Capacities

Citovat

HABA, Noam Y, Renana GROSS, Jiří NOVÁČEK, Hadassa SHAKED, Lukáš ŽÍDEK, Mira BARDA-SAAD and Jordan H CHILL. NMR Determines Transient Structure and Dynamics in the Disordered C-Terminal Domain of WASp Interacting Protein. Biophysical Journal. CAMBRIDGE: CELL PRESS, 2013, vol. 105, No 2, p. 481-493. ISSN 0006-3495. Available from: https://dx.doi.org/10.1016/j.bpj.2013.05.046.

@article{1123257,
   author = {Haba, Noam Y and Gross, Renana and Nováček, Jiří and Shaked, Hadassa and Žídek, Lukáš and BardaandSaad, Mira and Chill, Jordan H},
   article_location = {CAMBRIDGE},
   article_number = {2},
   doi = {http://dx.doi.org/10.1016/j.bpj.2013.05.046},
   keywords = {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},
   language = {eng},
   issn = {0006-3495},
   journal = {Biophysical Journal},
   title = {NMR Determines Transient Structure and Dynamics in the Disordered C-Terminal Domain of WASp Interacting Protein},
   url = {http://www.ncbi.nlm.nih.gov/pubmed/23870269},
   volume = {105},
   year = {2013}
}

TY  - JOUR
ID  - 1123257
AU  - Haba, Noam Y - Gross, Renana - Nováček, Jiří - Shaked, Hadassa - Žídek, Lukáš - Barda-Saad, Mira - Chill, Jordan H
PY  - 2013
TI  - NMR Determines Transient Structure and Dynamics in the Disordered C-Terminal Domain of WASp Interacting Protein
JF  - Biophysical Journal
VL  - 105
IS  - 2
SP  - 481-493
EP  - 481-493
PB  - CELL PRESS
SN  - 00063495
KW  - WISKOTT-ALDRICH-SYNDROME
KW  - NUCLEAR-MAGNETIC-RESONANCE
KW  - SEQUENCE-SPECIFIC ASSIGNMENT
KW  - PROLINE-RICH REGIONS
KW  - 8 M UREA
KW  - CHEMICAL-SHIFTS
KW  - ALPHA-SYNUCLEIN
KW  - N-WASP
KW  - ACTIN POLYMERIZATION
KW  - POLYPROLINE-II
UR  - http://www.ncbi.nlm.nih.gov/pubmed/23870269
L2  - http://www.ncbi.nlm.nih.gov/pubmed/23870269
N2  - 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.
ER  -

HABA, Noam Y, Renana GROSS, Jiří NOVÁČEK, Hadassa SHAKED, Lukáš ŽÍDEK, Mira BARDA-SAAD and Jordan H CHILL. NMR Determines Transient Structure and Dynamics in the Disordered C-Terminal Domain of WASp Interacting Protein. \textit{Biophysical Journal}. CAMBRIDGE: CELL PRESS, 2013, vol.~105, No~2, p.~481-493. ISSN~0006-3495. Available from: https://dx.doi.org/10.1016/j.bpj.2013.05.046.

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