Weak protein–cationic co-ion interactions addressed by X-ray crystallography and mass spectrometry
Autor: | Philippe Benas, Laurent Legrand, Anne Regazzetti, Franck Brachet, Nicolas Auzeil, Madeleine Riès-Kautt |
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Přispěvatelé: | Laboratoire de cristallographie et RMN biologiques (LCRB - UMR 8015), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie et Toxicologie Analytique et Cellulaire (EA 4463), Université Paris Descartes - Paris 5 (UPD5), Institut des Nanosciences de Paris (INSP), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Laboratoire de cristallographie et RMN biologiques ( LCRB - UMR 8015 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Chimie et Toxicologie Analytique et Cellulaire ( EA 4463 ), Université Paris Descartes - Paris 5 ( UPD5 ), Institut des Nanosciences de Paris ( INSP ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de biologie et chimie des protéines [Lyon] ( IBCP ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique ( CNRS ), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5) |
Jazyk: | angličtina |
Rok vydání: | 2014 |
Předmět: |
Spectrometry
Mass Electrospray Ionization Hofmeister series Electrospray ionization Analytical chemistry Salt (chemistry) 010402 general chemistry Mass spectrometry Crystallography X-Ray 01 natural sciences 03 medical and health sciences Structural Biology Cations Binding site 030304 developmental biology chemistry.chemical_classification [PHYS]Physics [physics] 0303 health sciences [ PHYS ] Physics [physics] Proteins General Medicine 0104 chemical sciences Crystallography chemistry X-ray crystallography Titration Adsorption Protein crystallization |
Zdroj: | Acta crystallographica Section D : Structural biology [1993-...] Acta crystallographica Section D : Structural biology [1993-..], 2014, 70 (8), pp.2217-2231. ⟨10.1107/S1399004714011304⟩ Acta Crystallographica Section D: Biological Crystallography Acta Crystallographica Section D: Biological Crystallography, International Union of Crystallography, 2014, 70 (8), pp.2217-2231. ⟨10.1107/S1399004714011304⟩ Acta Crystallographica Section D: Biological Crystallography, International Union of Crystallography, 2014, 70 (8), pp.2217-2231. 〈10.1107/S1399004714011304〉 |
ISSN: | 2059-7983 0907-4449 |
Popis: | The adsorption of Rb+, Cs+, Mn2+, Co2+and Yb3+onto the positively charged hen egg-white lysozyme (HEWL) has been investigated by solving 13 X-ray structures of HEWL crystallized with their chlorides and by applying electrospray ionization mass spectrometry (ESI-MS) first to dissolved protein crystals and then to the protein in buffered salt solutions. The number of bound cations follows the order Cs+< Mn2+≃ Co2+< Yb3+at 293 K. HEWL binds less Rb+(qtot= 0.7) than Cs+(qtot= 3.9) at 100 K. Crystal flash-cooling drastically increases the binding of Cs+, but poorly affects that of Yb3+, suggesting different interactions. The addition of glycerol increases the number of bound Yb3+cations, but only slightly increases that of Rb+. HEWL titrations with the same chlorides, followed by ESI-MS analysis, show that only about 10% of HEWL binds Cs+and about 40% binds 1–2 Yb3+cations, while the highest binding reaches 60–70% for protein binding 1–3 Mn2+or Co2+cations. The binding sites identified by X-ray crystallography show that the monovalent Rb+and Cs+preferentially bind to carbonyl groups, whereas the multivalent Mn2+, Co2+and Yb3+interact with carboxylic groups. This work elucidates the basis of the effect of the Hofmeister cation series on protein solubility. |
Databáze: | OpenAIRE |
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