title: Phosphorylation of the regulatory domain of human tyrosine hydroxylase 1 monitored using non-uniformly sampled NMR.
abstract: Human tyrosine hydroxylase 1 (hTH1) activity is regulated by phosphorylation of its regulatory domain (RD-hTH1) and by an interaction with the 14-3-3 protein. The RD-hTH1 is composed of a structured region (66-169) preceded by an intrinsically disordered protein region (IDP, hTH1_65) containing two phosphorylation sites (S19 and S40) which are highly relevant for its increase in activity. The NMR signals of the IDP region in the non-phosphorylated, singly phosphorylated (pS40) and doubly phosphorylated states (pS19_pS40) were assigned by non-uniformly sampled spectra with increased dimensionality (5D). The structural changes induced by phosphorylation were analyzed by means of secondary structure propensities. The phosphorylation kinetics of the S40 and S19 by kinases PKA and PRAK respectively were monitored by non-uniformly sampled time-resolved NMR spectroscopy followed by their quantitative analysis.
authors.affiliation: CEITEC MU, Masaryk University, Kamenice 753/5, 625 00 Brno, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Louša
authors.affiliation: CEITEC MU, Masaryk University, Kamenice 753/5, 625 00 Brno, Czech Republic.
authors.forename: Hana
authors.initials: H
authors.lastname: Nedozrálová
authors.affiliation: CEITEC MU, Masaryk University, Kamenice 753/5, 625 00 Brno, Czech Republic.
authors.forename: Erik
authors.initials: E
authors.lastname: Župa
authors.affiliation: CEITEC MU, Masaryk University, Kamenice 753/5, 625 00 Brno, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Nováček
authors.affiliation: CEITEC MU, Masaryk University, Kamenice 753/5, 625 00 Brno, Czech Republic. Electronic address: jozef.hritz@ceitec.muni.cz.
authors.forename: Jozef
authors.initials: J
authors.lastname: Hritz
mesh_terms: D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D010766:Phosphorylation; D000072417:Protein Domains; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D017433:Protein Structure, Secondary; D014446:Tyrosine 3-Monooxygenase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; C111049:MAP-kinase-activated kinase 5; D014446:Tyrosine 3-Monooxygenase; D017346:Protein Serine-Threonine Kinases; D017868:Cyclic AMP-Dependent Protein Kinases
keywords: Human tyrosine hydroxylase; IDP; Kinetics; NMR; Non-uniform sampling; Phosphorylation; SSP; Time-resolved NMR