5OKF

CH1 chimera of human 14-3-3 sigma with the HSPB6 phosphopeptide in a conformation with self-bound phosphopeptides

Summary for 5OKF

• Entry DOI: 10.2210/pdb5okf/pdb
• Descriptor: 14-3-3 protein sigma,Heat shock protein beta-6, CADMIUM ION (3 entities in total)
• Functional Keywords: 14-3-3 proteins, protein chimera, phosphopeptide-binding, signaling protein
• Biological source: Homo sapiens (Human); More
• Cellular location: Cytoplasm : O14558
• Total number of polymer chains: 4
• Total formula weight: 111946.56

Authors

Sluchanko, N.N.,Tugaeva, K.V.,Greive, S.J.,Antson, A.A. (deposition date: 2017-07-25, release date: 2017-10-11, Last modification date: 2024-10-16)

Primary citation

Sluchanko, N.N.,Tugaeva, K.V.,Greive, S.J.,Antson, A.A.
Chimeric 14-3-3 proteins for unraveling interactions with intrinsically disordered partners.
Sci Rep, 7:12014-12014, 2017

PubMed Abstract: In eukaryotes, several "hub" proteins integrate signals from different interacting partners that bind through intrinsically disordered regions. The 14-3-3 protein hub, which plays wide-ranging roles in cellular processes, has been linked to numerous human disorders and is a promising target for therapeutic intervention. Partner proteins usually bind via insertion of a phosphopeptide into an amphipathic groove of 14-3-3. Structural plasticity in the groove generates promiscuity allowing accommodation of hundreds of different partners. So far, accurate structural information has been derived for only a few 14-3-3 complexes with phosphopeptide-containing proteins and a variety of complexes with short synthetic peptides. To further advance structural studies, here we propose a novel approach based on fusing 14-3-3 proteins with the target partner peptide sequences. Such chimeric proteins are easy to design, express, purify and crystallize. Peptide attachment to the C terminus of 14-3-3 via an optimal linker allows its phosphorylation by protein kinase A during bacterial co-expression and subsequent binding at the amphipathic groove. Crystal structures of 14-3-3 chimeras with three different peptides provide detailed structural information on peptide-14-3-3 interactions. This simple but powerful approach, employing chimeric proteins, can reinvigorate studies of 14-3-3/phosphoprotein assemblies, including those with challenging low-affinity partners, and may facilitate the design of novel biosensors.

PubMed: 28931924
DOI: 10.1038/s41598-017-12214-9

Experimental method

X-RAY DIFFRACTION (3.2 Å)