Next Article in Journal
Genetic Isolation among the Northwestern, Southwestern and Central-Eastern Indian Ocean Populations of the Pronghorn Spiny Lobster Panulirus penicillatus
Next Article in Special Issue
Mechanisms of Enzyme-Catalyzed Reduction of Two Carcinogenic Nitro-Aromatics, 3-Nitrobenzanthrone and Aristolochic Acid I: Experimental and Theoretical Approaches
Previous Article in Journal
A Simple Three-Step Method for Design and Affinity Testing of New Antisense Peptides: An Example of Erythropoietin
Previous Article in Special Issue
Application of Computational Methods for the Design of BACE-1 Inhibitors: Validation of in Silico Modelling
Int. J. Mol. Sci. 2014, 15(6), 9224-9241; doi:10.3390/ijms15069224
Article

The Application of an Emerging Technique for Protein–Protein Interaction Interface Mapping: The Combination of Photo-Initiated Cross-Linking Protein Nanoprobes with Mass Spectrometry

1,2
,
1,2
,
1,2
,
2
,
2
 and
1,2,*
1 Institute of Microbiology v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, CZ-14220 Prague 4, Czech Republic 2 Department of Biochemistry, Faculty of Science, Charles University, Hlavova 2030, CZ-12843 Prague 2, Czech Republic
* Author to whom correspondence should be addressed.
Received: 17 January 2014 / Revised: 6 May 2014 / Accepted: 9 May 2014 / Published: 26 May 2014
View Full-Text   |   Download PDF [1150 KB, 19 June 2014; original version 19 June 2014]   |  

Abstract

Protein–protein interaction was investigated using a protein nanoprobe capable of photo-initiated cross-linking in combination with high-resolution and tandem mass spectrometry. This emerging experimental approach introduces photo-analogs of amino acids within a protein sequence during its recombinant expression, preserves native protein structure and is suitable for mapping the contact between two proteins. The contact surface regions involved in the well-characterized interaction between two molecules of human 14-3-3ζ regulatory protein were used as a model. The employed photo-initiated cross-linking techniques extend the number of residues shown to be within interaction distance in the contact surface of the 14-3-3ζ dimer (Gln8–Met78). The results of this study are in agreement with our previously published data from molecular dynamic calculations based on high-resolution chemical cross-linking data and Hydrogen/Deuterium exchange mass spectrometry. The observed contact is also in accord with the 14-3-3ζ X-ray crystal structure (PDB 3dhr). The results of the present work are relevant to the structural biology of transient interaction in the 14-3-3ζ protein, and demonstrate the ability of the chosen methodology (the combination of photo-initiated cross-linking protein nanoprobes and mass spectrometry analysis) to map the protein-protein interface or regions with a flexible structure.
Keywords: 14-3-3ζ homodimer; protein-protein interaction; photo cross-linking; protein nanoprobe; mass spectrometry 14-3-3ζ homodimer; protein-protein interaction; photo cross-linking; protein nanoprobe; mass spectrometry
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
SciFeed

Share & Cite This Article

Further Mendeley | CiteULike
Export to BibTeX |
EndNote |
RIS
MDPI and ACS Style

Ptáčková, R.; Ječmen, T.; Novák, P.; Hudeček, J.; Stiborová, M.; Šulc, M. The Application of an Emerging Technique for Protein–Protein Interaction Interface Mapping: The Combination of Photo-Initiated Cross-Linking Protein Nanoprobes with Mass Spectrometry. Int. J. Mol. Sci. 2014, 15, 9224-9241.

View more citation formats

Related Articles

Article Metrics

For more information on the journal, click here

Comments

[Return to top]
Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert