Structure, Dynamics and Thermodynamics of the Human Centrin 2/hSfi1 Complex
| Autor: | Geoffrey Bodenhausen, Liliane Assairi, Liliane Mouawad, Constantin T. Craescu, Daniel Abergel, Fatiha Kateb, Yves Blouquit, Juan Martinez-Sanz |
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| Přispěvatelé: | Institut des sciences du végétal (ISV), Centre National de la Recherche Scientifique (CNRS), Biomolécules : synthèse, structure et mode d'action (UMR 8642) (BIOSYMA), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut Curie [Paris], Imagerie intégrative de la molécule à l'organisme, Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut des sciences et d'ingénierie chimiques (ISIC), Ecole Polytechnique Fédérale de Lausanne (EPFL), Université Pierre et Marie Curie - Paris 6 (UPMC), Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), École normale supérieure - Paris (ENS-PSL) |
| Rok vydání: | 2010 |
| Předmět: |
Models
Molecular Magnetic Resonance Spectroscopy human centrin 2 Cell Cycle Proteins Peptide Crystallography X-Ray Protein Structure Secondary 0302 clinical medicine Structural Biology Sfi1 chemistry.chemical_classification 0303 health sciences [CHIM.ORGA]Chemical Sciences/Organic chemistry Hydrogen bond Backbone Dynamics Solutions Biochemistry Thermodynamics Molecular-Cloning Two-dimensional nuclear magnetic resonance spectroscopy Heteronuclear single quantum coherence spectroscopy Protein Binding Repetitive Sequences Amino Acid DNA repair Quantum Nmr-Spectroscopy Excision-Repair Molecular Sequence Data Calorimetry Biology Group-C Protein 03 medical and health sciences multiple-quantum relaxation Chlamydomonas-Reinhardtii Mitotic Spindle Poles Humans Chemical-Shift Modulations Amino Acid Sequence Molecular Biology 030304 developmental biology Binding Sites Cross-Correlated Relaxation Calcium-Binding Proteins Itc Nmr chemistry Centrosome Centrin Pole Body Duplication Biophysics Calcium Peptides 030217 neurology & neurosurgery Nucleotide excision repair |
| Zdroj: | Journal of Molecular Biology Journal of Molecular Biology, Elsevier, 2010, 395 (1), pp.191-204. ⟨10.1016/j.jmb.2009.10.041⟩ Journal of Molecular Biology, Elsevier, 2010, 395 ((1)), pp.191-204. ⟨10.1016/j.jmb.2009.10.041⟩ Journal of Molecular Biology, 2010, 395 (1), pp.191-204. ⟨10.1016/j.jmb.2009.10.041⟩ |
| ISSN: | 0022-2836 1089-8638 |
| DOI: | 10.1016/j.jmb.2009.10.041 |
| Popis: | Centrin, an EF-hand calcium-binding protein, has been shown to be involved in the duplication of centrosomes, and Sfi1 (Suppressor of fermentation-induced loss of stress resistance protein 1) is one of its centrosomal targets. There are three isoforms of human centrin, but here we only considered centrin 2 (HsCen2). This protein has the ability to bind to any of the similar to 25 repeats of human Sfi1 (hSfi1) with more or less affinity. In this study, we mainly focused on the 17th repeat (R17-hSfi1-20), which presents the highest level of similarity with a well-studied 17-residue peptide (P17-XPC) from human xeroderma pigmentosum complementation group C protein, another centrin target for DNA repair. The only known structure of HsCen2 was resolved in complex with P17-XPC. The 20-residue peptide R17-hSfi1-20 exhibits the motif L8L4W1, which is the reverse of the XPC motif, W1L4L8. Consequently, the dipole of the helix formed by this motif has a reverse orientation. We wished to ascertain the impact of this reversal on the structure, dynamics and affinity of centrin. To address this question, we determined the structure of C-HsCen2 [the C-terminal domain of HsCen2 (T94-Y172)] in complex with R17-hSfi1-20 and monitored its dynamics by NMR, after having verified that the N-terminal domain of HsCen2 does not interact with the peptide The structure shows that the binding mode is similar to that of P17-XPC. However, we observed a 2 -angstrom translation of the R17-hSfi1-20 helix along its axis, inducing less anchorage in the protein and the disruption of a hydrogen bond between a tryptophan residue in the peptide and a well-conserved nearby glutamate in C+HsCen2. NMR dynamic studies of the complex strongly suggested the existence of an unusual calcium secondary binding mode in calcium-binding loop III made possible by the uncommon residue composition of this loop. The secondary metal site is only populated at high calcium concentration and depends on the type of bound ligand. (C) 2009 Elsevier Ltd. All rights reserved |
| Databáze: | OpenAIRE |
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