Structure of the NPr:EINNtr Complex: Mechanism for Specificity in Paralogous Phosphotransferase Systems

Madeleine Strickland, Ann Marie Stanley, Guangshun Wang, Istvan Botos, Charles D. Schwieters, Susan K. Buchanan, Alan Peterkofsky, Nico Tjandra

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Paralogous enzymes arise from gene duplication events that confer a novel function, although it is unclear how cross-reaction between the original and duplicate protein interaction network is minimized. We investigated HPr:EIsugar and NPr:EINtr, the initial complexes of paralogous phosphorylation cascades involved in sugar import and nitrogen regulation in bacteria, respectively. Although the HPr:EIsugar interaction has been well characterized, involving multiple complexes and transient interactions, the exact nature of the NPr:EINtr complex was unknown. We set out to identify the key features of the interaction by performing binding assays and elucidating the structure of NPr in complex with the phosphorylation domain of EINtr (EINNtr), using a hybrid approach involving X-ray, homology, and sparse nuclear magnetic resonance. We found that the overall fold and active-site structure of the two complexes are conserved in order to maintain productive phosphorylation, however, the interface surface potential differs between the two complexes, which prevents cross-reaction.

Original languageEnglish (US)
Pages (from-to)2127-2137
Number of pages11
JournalStructure
Volume24
Issue number12
DOIs
StatePublished - Dec 6 2016

Fingerprint

Phosphotransferases
Cross Reactions
Phosphorylation
Protein Interaction Maps
Gene Duplication
Catalytic Domain
Magnetic Resonance Spectroscopy
Nitrogen
X-Rays
Bacteria
Enzymes

Keywords

  • Enzyme I
  • NPr
  • X-ray crystallography
  • nuclear magnetic resonance
  • phosphotransferase system
  • pseudocontact shifts
  • residual dipolar couplings
  • small-angle X-ray scattering
  • specificity
  • surface potential

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Strickland, M., Stanley, A. M., Wang, G., Botos, I., Schwieters, C. D., Buchanan, S. K., ... Tjandra, N. (2016). Structure of the NPr:EINNtr Complex: Mechanism for Specificity in Paralogous Phosphotransferase Systems. Structure, 24(12), 2127-2137. https://doi.org/10.1016/j.str.2016.10.007

Structure of the NPr:EINNtr Complex : Mechanism for Specificity in Paralogous Phosphotransferase Systems. / Strickland, Madeleine; Stanley, Ann Marie; Wang, Guangshun; Botos, Istvan; Schwieters, Charles D.; Buchanan, Susan K.; Peterkofsky, Alan; Tjandra, Nico.

In: Structure, Vol. 24, No. 12, 06.12.2016, p. 2127-2137.

Research output: Contribution to journalArticle

Strickland, M, Stanley, AM, Wang, G, Botos, I, Schwieters, CD, Buchanan, SK, Peterkofsky, A & Tjandra, N 2016, 'Structure of the NPr:EINNtr Complex: Mechanism for Specificity in Paralogous Phosphotransferase Systems', Structure, vol. 24, no. 12, pp. 2127-2137. https://doi.org/10.1016/j.str.2016.10.007
Strickland, Madeleine ; Stanley, Ann Marie ; Wang, Guangshun ; Botos, Istvan ; Schwieters, Charles D. ; Buchanan, Susan K. ; Peterkofsky, Alan ; Tjandra, Nico. / Structure of the NPr:EINNtr Complex : Mechanism for Specificity in Paralogous Phosphotransferase Systems. In: Structure. 2016 ; Vol. 24, No. 12. pp. 2127-2137.
@article{b2db49073f1d4318bb19f8d3cdf12355,
title = "Structure of the NPr:EINNtr Complex: Mechanism for Specificity in Paralogous Phosphotransferase Systems",
abstract = "Paralogous enzymes arise from gene duplication events that confer a novel function, although it is unclear how cross-reaction between the original and duplicate protein interaction network is minimized. We investigated HPr:EIsugar and NPr:EINtr, the initial complexes of paralogous phosphorylation cascades involved in sugar import and nitrogen regulation in bacteria, respectively. Although the HPr:EIsugar interaction has been well characterized, involving multiple complexes and transient interactions, the exact nature of the NPr:EINtr complex was unknown. We set out to identify the key features of the interaction by performing binding assays and elucidating the structure of NPr in complex with the phosphorylation domain of EINtr (EINNtr), using a hybrid approach involving X-ray, homology, and sparse nuclear magnetic resonance. We found that the overall fold and active-site structure of the two complexes are conserved in order to maintain productive phosphorylation, however, the interface surface potential differs between the two complexes, which prevents cross-reaction.",
keywords = "Enzyme I, NPr, X-ray crystallography, nuclear magnetic resonance, phosphotransferase system, pseudocontact shifts, residual dipolar couplings, small-angle X-ray scattering, specificity, surface potential",
author = "Madeleine Strickland and Stanley, {Ann Marie} and Guangshun Wang and Istvan Botos and Schwieters, {Charles D.} and Buchanan, {Susan K.} and Alan Peterkofsky and Nico Tjandra",
year = "2016",
month = "12",
day = "6",
doi = "10.1016/j.str.2016.10.007",
language = "English (US)",
volume = "24",
pages = "2127--2137",
journal = "Structure with Folding & design",
issn = "0969-2126",
publisher = "Cell Press",
number = "12",

}

TY - JOUR

T1 - Structure of the NPr:EINNtr Complex

T2 - Mechanism for Specificity in Paralogous Phosphotransferase Systems

AU - Strickland, Madeleine

AU - Stanley, Ann Marie

AU - Wang, Guangshun

AU - Botos, Istvan

AU - Schwieters, Charles D.

AU - Buchanan, Susan K.

AU - Peterkofsky, Alan

AU - Tjandra, Nico

PY - 2016/12/6

Y1 - 2016/12/6

N2 - Paralogous enzymes arise from gene duplication events that confer a novel function, although it is unclear how cross-reaction between the original and duplicate protein interaction network is minimized. We investigated HPr:EIsugar and NPr:EINtr, the initial complexes of paralogous phosphorylation cascades involved in sugar import and nitrogen regulation in bacteria, respectively. Although the HPr:EIsugar interaction has been well characterized, involving multiple complexes and transient interactions, the exact nature of the NPr:EINtr complex was unknown. We set out to identify the key features of the interaction by performing binding assays and elucidating the structure of NPr in complex with the phosphorylation domain of EINtr (EINNtr), using a hybrid approach involving X-ray, homology, and sparse nuclear magnetic resonance. We found that the overall fold and active-site structure of the two complexes are conserved in order to maintain productive phosphorylation, however, the interface surface potential differs between the two complexes, which prevents cross-reaction.

AB - Paralogous enzymes arise from gene duplication events that confer a novel function, although it is unclear how cross-reaction between the original and duplicate protein interaction network is minimized. We investigated HPr:EIsugar and NPr:EINtr, the initial complexes of paralogous phosphorylation cascades involved in sugar import and nitrogen regulation in bacteria, respectively. Although the HPr:EIsugar interaction has been well characterized, involving multiple complexes and transient interactions, the exact nature of the NPr:EINtr complex was unknown. We set out to identify the key features of the interaction by performing binding assays and elucidating the structure of NPr in complex with the phosphorylation domain of EINtr (EINNtr), using a hybrid approach involving X-ray, homology, and sparse nuclear magnetic resonance. We found that the overall fold and active-site structure of the two complexes are conserved in order to maintain productive phosphorylation, however, the interface surface potential differs between the two complexes, which prevents cross-reaction.

KW - Enzyme I

KW - NPr

KW - X-ray crystallography

KW - nuclear magnetic resonance

KW - phosphotransferase system

KW - pseudocontact shifts

KW - residual dipolar couplings

KW - small-angle X-ray scattering

KW - specificity

KW - surface potential

UR - http://www.scopus.com/inward/record.url?scp=85002842137&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85002842137&partnerID=8YFLogxK

U2 - 10.1016/j.str.2016.10.007

DO - 10.1016/j.str.2016.10.007

M3 - Article

C2 - 27839951

AN - SCOPUS:85002842137

VL - 24

SP - 2127

EP - 2137

JO - Structure with Folding & design

JF - Structure with Folding & design

SN - 0969-2126

IS - 12

ER -