21920446 OR Structural basis of 14-3-3 protein functions
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The 14-3-3 proteins, a family of conserved regulatory molecules, participate in a wide range of cellular processes through binding interactions with hundreds of structurally and functionally diverse proteins. Several distinct mechanisms of the 14-3-3 protein function were described, including conformational modulation of the bound protein, masking of its sequence-specific or structural features, and scaffolding that facilitates interaction between two simultaneously bound proteins. Details of these functional modes, especially from the structural point of view, still remain mostly elusive. This review gives an overview of the current knowledge concerning the structure of 14-3-3 proteins and their complexes as well as the insights it provides into the mechanisms of their functions. We discuss structural basis of target recognition by 14-3-3 proteins, common structural features of their complexes and known mechanisms of 14-3-3 protein-dependent regulations.
- MeSH
- DNA vazebné proteiny MeSH
- Eukaryota metabolismus MeSH
- fosforylace MeSH
- genetická variace MeSH
- interakční proteinové domény a motivy MeSH
- konformace proteinů MeSH
- lidé MeSH
- molekulární modely MeSH
- protein - isoformy chemie metabolismus MeSH
- proteiny 14-3-3 chemie genetika metabolismus MeSH
- terciární struktura proteinů MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Regulator of G protein signaling (RGS) proteins function as GTPase-activating proteins for the α-subunit of heterotrimeric G proteins. The function of certain RGS proteins is negatively regulated by 14-3-3 proteins, a family of highly conserved regulatory molecules expressed in all eukaryotes. In this study, we provide a structural mechanism for 14-3-3-dependent inhibition of RGS3-Gα interaction. We have used small angle x-ray scattering, hydrogen/deuterium exchange kinetics, and Förster resonance energy transfer measurements to determine the low-resolution solution structure of the 14-3-3ζ·RGS3 complex. The structure shows the RGS domain of RGS3 bound to the 14-3-3ζ dimer in an as-yet-unrecognized manner interacting with less conserved regions on the outer surface of the 14-3-3 dimer outside its central channel. Our results suggest that the 14-3-3 protein binding affects the structure of the Gα interaction portion of RGS3 as well as sterically blocks the interaction between the RGS domain and the Gα subunit of heterotrimeric G proteins.
- MeSH
- cirkulární dichroismus MeSH
- fosforylace MeSH
- hmotnostní spektrometrie MeSH
- lidé MeSH
- maloúhlový rozptyl MeSH
- proteiny 14-3-3 chemie genetika metabolismus MeSH
- proteiny aktivující GTPasu chemie genetika metabolismus MeSH
- proteiny vázající GTP chemie genetika metabolismus MeSH
- rezonanční přenos fluorescenční energie MeSH
- sekundární struktura proteinů MeSH
- signální transdukce MeSH
- terciární struktura proteinů MeSH
- vazba proteinů MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
14-3-3 proteins are important dimeric scaffolds that regulate the function of hundreds of proteins in a phosphorylation-dependent manner. The SARS-CoV-2 nucleocapsid (N) protein forms a complex with human 14-3-3 proteins upon phosphorylation, which has also been described for other coronaviruses. Here, we report a high-resolution crystal structure of 14-3-3 bound to an N phosphopeptide bearing the phosphoserine 197 in the middle. The structure revealed two copies of the N phosphopeptide bound, each in the central binding groove of each 14-3-3 monomer. A complex network of hydrogen bonds and water bridges between the peptide and 14-3-3 was observed explaining the high affinity of the N protein for 14-3-3 proteins.
- MeSH
- COVID-19 MeSH
- fosfopeptidy chemie MeSH
- fosfoproteiny chemie MeSH
- koronavirové nukleokapsidové proteiny * chemie MeSH
- lidé MeSH
- proteiny 14-3-3 * chemie MeSH
- SARS-CoV-2 * MeSH
- vazba proteinů MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Phosducin (Pdc) is a conserved phosphoprotein that, when unphosphorylated, binds with high affinity to the complex of βγ-subunits of G protein transducin (Gtβγ). The ability of Pdc to bind to Gtβγ is inhibited through its phosphorylation at S54 and S73 within the N-terminal domain (Pdc-ND) followed by association with the scaffolding protein 14-3-3. However, the molecular basis for the 14-3-3-dependent inhibition of Pdc binding to Gtβγ is unclear. By using small-angle x-ray scattering, high-resolution NMR spectroscopy, and limited proteolysis coupled with mass spectrometry, we show that phosphorylated Pdc and 14-3-3 form a complex in which the Pdc-ND region 45-80, which forms a part of Pdc's Gtβγ binding surface and contains both phosphorylation sites, is restrained within the central channel of the 14-3-3 dimer, with both 14-3-3 binding motifs simultaneously participating in protein association. The N-terminal part of Pdc-ND is likely located outside the central channel of the 14-3-3 dimer, but Pdc residues 20-30, which are also involved in Gtβγ binding, are positioned close to the surface of the 14-3-3 dimer. The C-terminal domain of Pdc is located outside the central channel and its structure is unaffected by the complex formation. These results indicate that the 14-3-3 protein-mediated inhibition of Pdc binding to Gtβγ is based on steric occlusion of Pdc's Gtβγ binding surface.
- MeSH
- difrakce rentgenového záření MeSH
- fosfoproteiny antagonisté a inhibitory chemie MeSH
- fosforylace MeSH
- krysa rodu rattus MeSH
- maloúhlový rozptyl MeSH
- oční proteiny antagonisté a inhibitory chemie MeSH
- proteinové domény MeSH
- proteiny 14-3-3 chemie metabolismus MeSH
- proteiny vázající GTP - regulátory antagonisté a inhibitory chemie MeSH
- proteolýza MeSH
- protonová magnetická rezonanční spektroskopie MeSH
- sekundární struktura proteinů MeSH
- vazba proteinů MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The 14-3-3 proteins, a family of highly conserved scaffolding proteins ubiquitously expressed in all eukaryotic cells, interact with and regulate the function of several hundreds of partner proteins. Yeast neutral trehalases (Nth), enzymes responsible for the hydrolysis of trehalose to glucose, compared with trehalases from other organisms, possess distinct structure and regulation involving phosphorylation at multiple sites followed by binding to the 14-3-3 protein. Here we report the crystal structures of yeast Nth1 and its complex with Bmh1 (yeast 14-3-3 isoform), which, together with mutational and fluorescence studies, indicate that the binding of Nth1 by 14-3-3 triggers Nth1's activity by enabling the proper 3D configuration of Nth1's catalytic and calcium-binding domains relative to each other, thus stabilizing the flexible part of the active site required for catalysis. The presented structure of the Bmh1:Nth1 complex highlights the ability of 14-3-3 to modulate the structure of a multidomain binding partner and to function as an allosteric effector. Furthermore, comparison of the Bmh1:Nth1 complex structure with those of 14-3-3:serotonin N-acetyltransferase and 14-3-3:heat shock protein beta-6 complexes revealed similarities in the 3D structures of bound partner proteins, suggesting the highly conserved nature of 14-3-3 affects the structures of many client proteins.
- MeSH
- arylalkylamin-N-acetyltransferasa metabolismus MeSH
- chemické databáze * MeSH
- fosforylace MeSH
- glukosa metabolismus MeSH
- katalytická doména MeSH
- konformace proteinů MeSH
- krystalografie rentgenová MeSH
- molekulární modely MeSH
- proteinové domény MeSH
- proteiny 14-3-3 genetika metabolismus MeSH
- proteiny teplotního šoku chemie metabolismus MeSH
- Saccharomyces cerevisiae - proteiny chemie metabolismus MeSH
- Saccharomyces cerevisiae enzymologie genetika metabolismus MeSH
- trehalasa chemie metabolismus MeSH
- trehalosa metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Na+/H+ antiporters are involved in ensuring optimal intracellular concentrations of alkali-metal cations and protons in most organisms. In Saccharomyces cerevisiae, the plasma-membrane Na+, K+/H+ antiporter Nha1 mediates Na+ and K+ efflux, which is important for cell growth in the presence of salts. Nha1 belongs among housekeeping proteins and, due to its ability to export K+, it has many physiological functions. The Nha1 transport activity is regulated through its long, hydrophilic and unstructured C-terminus (554 of 985 aa). Although Nha1 has been previously shown to interact with the yeast 14-3-3 isoform (Bmh2), the binding site remains unknown. In this work, we identified the residues through which Nha1 interacts with the 14-3-3 protein. Biophysical characterization of the interaction between the C-terminal polypeptide of Nha1 and Bmh proteins in vitro revealed that the 14-3-3 protein binds to phosphorylated Ser481 of Nha1, and the crystal structure of the phosphopeptide containing Ser481 bound to Bmh1 provided the structural basis of this interaction. Our data indicate that 14-3-3 binding induces a disorder-to-order transition of the C-terminus of Nha1, and in vivo experiments showed that the mutation of Ser481 to Ala significantly increases cation efflux activity via Nha1, which renders cells sensitive to low K+ concentrations. Hence, 14-3-3 binding is apparently essential for the negative regulation of Nha1 activity, which should be low under standard growth conditions, when low amounts of toxic salts are present and yeast cells need to accumulate high amounts of K+.
... Hrazdíra) 1 -- 2 The structure of living matter (V. ... ... Mornstein) 3 -- 2.1 A recapitulation of quantum and nuclear physics 3 -- 2.1.1 The sub-molecular structure ... ... of matter 3 -- 2.1.1.1 The four fundamental interactions 3 -- 2.1.1.2 Elementary particles of matter ... ... of water in organisms 26 -- 2.2.3 Biopolymers and their structure 27 -- 2.2.3.1 Structure of nucleic ... ... acids 27 -- 2.2.3.2 Protein structures and their modifications 28 -- 2.2.3.3 Overview of the methods ...
1. vyd. viii, 317 s. : il. ; 30 cm
... Hrazdíra) 9 -- 2 The structure of living matter (V. ... ... Mornstein) 11 -- 2.1 A recapitulation of quantum and nuclear physics 11 -- 2.1.1 The sub-molecular structure ... ... of water in organisms 34 -- 2.2.3 Biopolymers and their structure 35 -- 2.2.3.1 Structure of nucleic ... ... acids 35 -- 2.2.3.2 Protein structures and their modifications 36 -- 2.2.3.3 Overview of the methods ... ... basis of invasive therapeutic methods and equipment assisting or replacing organ functions (I. ...
2nd rev. ed. 325 s. : il., tab. ; 30 cm
- Konspekt
- Biochemie. Molekulární biologie. Biofyzika
- NLK Obory
- fyzika, biofyzika
- NLK Publikační typ
- učebnice vysokých škol
