Human Nek6 is a monomeric mostly globular kinase with an unfolded short N-terminal domain

The NIMA-related kinases (Neks) are widespread among eukaryotes. In mammalians they represent an evolutionarily conserved family of 11 serine/threonine kinases, with 40-45% amino acid sequence identity to the Aspergillus nidulans mitotic regulator NIMA within their catalytic domains. Neks have cell...

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Bibliographic Details
Published in:BMC structural biology 2011-02, Vol.11 (1), p.12-12, Article 12
Main Authors: Meirelles, Gabriela V, Silva, Júlio C, Mendonça, Yuri de A, Ramos, Carlos Hi, Torriani, Iris L, Kobarg, Jörg
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Cancer
Chromatography, Gel
Cyclin-dependent kinases
Experiments
Gene expression
Genetic aspects
Humans
Kinases
Light
Models, Molecular
Molecular Sequence Data
NIMA-Related Kinases
Phosphorylation
Phosphotransferases
Physiological aspects
Protein Serine-Threonine Kinases - chemistry
Protein Serine-Threonine Kinases - metabolism
Protein Structure, Secondary
Protein Structure, Tertiary
Protein Unfolding
Proteins
Regulation
Scattering, Small Angle
Software
Solutions
X-Ray Diffraction
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Summary:The NIMA-related kinases (Neks) are widespread among eukaryotes. In mammalians they represent an evolutionarily conserved family of 11 serine/threonine kinases, with 40-45% amino acid sequence identity to the Aspergillus nidulans mitotic regulator NIMA within their catalytic domains. Neks have cell cycle-related functions and were recently described as related to pathologies, particularly cancer, consisting in potential chemotherapeutic targets. Human Nek6, -7 and -9 are involved in the control of mitotic spindle formation, acting together in a mitotic kinase cascade, but their mechanism of regulation remain elusive. In this study we performed a biophysical and structural characterization of human Nek6 with the aim of obtaining its low resolution and homology models. SAXS experiments showed that hNek6 is a monomer of a mostly globular, though slightly elongated shape. Comparative molecular modeling together with disorder prediction analysis also revealed a flexible disordered N-terminal domain for hNek6, which we found to be important to mediate interactions with diverse partners. SEC-MALS experiments showed that hNek6 conformation is dependent on its activation/phosphorylation status, a higher phosphorylation degree corresponding to a bigger Stokes radius. Circular dichroism spectroscopy confirmed our in silico predictions of secondary structure content and thermal stability shift assays revealed a slightly higher stability of wild-type hNek6 compared to the activation loop mutant hNek6(S206A). Our data present the first low resolution 3D structure of hNek6 protein in solution. SAXS, comparative modeling and SEC-MALS analysis revealed that hNek6 is a monomeric kinase of slightly elongated shape and a short unfolded N-terminal domain.
ISSN:1472-6807
1472-6807
DOI:10.1186/1472-6807-11-12