Dissection of Binding between a Phosphorylated Tyrosine Hydroxylase Peptide and 14-3-3zéta: A Complex Story Elucidated by NMR

HRITZ, Jozef, In-Ja L. BYEON, Troyi KRZYSIAK, Aurora MARTINEZ, Vladimír SKLENÁŘ and Angela M. GRONENBORN. Dissection of Binding between a Phosphorylated Tyrosine Hydroxylase Peptide and 14-3-3zéta: A Complex Story Elucidated by NMR. Biophysical Journal. New York, USA: Cell Press, 2014, vol. 107, No 9, p. 2185-2194. ISSN 0006-3495. Available from: https://dx.doi.org/10.1016/j.bpj.2014.08.039.

Basic information

• Original name: Dissection of Binding between a Phosphorylated Tyrosine Hydroxylase Peptide and 14-3-3zéta: A Complex Story Elucidated by NMR
• Authors: HRITZ, Jozef (703 Slovakia, guarantor, belonging to the institution), In-Ja L. BYEON (840 United States of America), Troyi KRZYSIAK (840 United States of America), Aurora MARTINEZ (578 Norway), Vladimír SKLENÁŘ (203 Czech Republic, belonging to the institution) and Angela M. GRONENBORN (840 United States of America).
• Edition: Biophysical Journal, New York, USA, Cell Press, 2014, 0006-3495.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10610 Biophysics
• Country of publisher: United States of America
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• Impact factor: Impact factor: 3.972
• RIV identification code: RIV/00216224:14740/14:00077266
• Organization unit: Central European Institute of Technology
• Doi: http://dx.doi.org/10.1016/j.bpj.2014.08.039
• UT WoS: 000344232500020
• Keywords in English: STRUCTURAL BASIS; PROTEINS; DOMAIN; SITES; GENE
• Tags: kontrola MP, MP, rivok
• Tags: International impact, Reviewed

Abstract

Human tyrosine hydroxylase activity is regulated by phosphorylation of its N-terminus and by an interaction with the modulator 14-3-3 proteins. We investigated the binding of singly or doubly phosphorylated and thiophosphorylated peptides, comprising the first 50 amino acids of human tyrosine hydroxylase, isoform 1 (hTH1), that contain the critical interaction domain, to 14-3-3zéta, by 31P NMR. Single phosphorylation at S19 generates a high affinity 14-3-3zéta binding epitope, whereas singly S40-phosphorylated peptide interacts with 14-3-3zéta one order-of-magnitude weaker than the S19-phosphorylated peptide. Analysis of the binding data revealed that the 14-3-3zéta dimer and the S19- and S40-doubly phosphorylated peptide interact in multiple ways, with three major complexes formed: 1), a single peptide bound to a 14-3-3zéta dimer via the S19 phosphate with the S40 phosphate occupying the other binding site; 2), a single peptide bound to a 14-3-3zéta dimer via the S19 phosphorous with the S40 free in solution; or 3), a 14-3-3zéta dimer with two peptides bound via the S19 phosphorous to each binding site. Our system and data provide information as to the possible mechanisms by which 14-3-3 can engage binding partners that possess two phosphorylation sites on flexible tails. Whether these will be realized in any particular interacting pair will naturally depend on the details of each system.