A Pozza, A Martel, M Moir, T A Darwish, K Wimalan, A Koutsioubas, S Combet, F Bonneté
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引用次数: 0
Abstract
In this study, we investigate the detergent-induced behavior of the integral membrane protein ShuA in solution, focusing on its interactions with octyl polyoxyethylene (OPOE) and n-dodecyl-β-D-maltoside (DDM). Using a combination of size-exclusion chromatography coupled with multi-angle light scattering (SEC-MALS) and small-angle scattering techniques (SAXS and SANS), we provide a detailed characterization of the protein-detergent complex (PDC) behavior under varying conditions. Our results reveal that ShuA remains monomeric in 1% OPOE, whereas in 0.5 mM DDM, it undergoes a reversible monomer/dimer equilibrium that shifts towards a monodisperse, monomeric state with increasing DDM concentration to 7.5 mM, highlighting the significant influence of detergent type and concentration on protein colloidal stability. These findings have direct implications for membrane protein purification and structural studies, particularly in crystallization and cryo-EM sample preparation. The study emphasizes the necessity of optimizing detergent conditions to ensure monodispersity and structural integrity, preventing detergent-induced artifacts that could affect structural interpretations. Importantly, our results highlight the power of the SEC-MALS technique in determining oligomeric or association equilibrium states, detecting weak intermolecular interactions often overlooked in conventional SEC, and achieving this even in the particularly complex case of MPs. By integrating advanced scattering techniques, this work contributes valuable insights into MP colloidal behavior, refining strategies for structural characterization and providing a framework for optimizing detergent conditions in biochemical and biophysical studies.
期刊介绍:
Protein Science, the flagship journal of The Protein Society, is a publication that focuses on advancing fundamental knowledge in the field of protein molecules. The journal welcomes original reports and review articles that contribute to our understanding of protein function, structure, folding, design, and evolution.
Additionally, Protein Science encourages papers that explore the applications of protein science in various areas such as therapeutics, protein-based biomaterials, bionanotechnology, synthetic biology, and bioelectronics.
The journal accepts manuscript submissions in any suitable format for review, with the requirement of converting the manuscript to journal-style format only upon acceptance for publication.
Protein Science is indexed and abstracted in numerous databases, including the Agricultural & Environmental Science Database (ProQuest), Biological Science Database (ProQuest), CAS: Chemical Abstracts Service (ACS), Embase (Elsevier), Health & Medical Collection (ProQuest), Health Research Premium Collection (ProQuest), Materials Science & Engineering Database (ProQuest), MEDLINE/PubMed (NLM), Natural Science Collection (ProQuest), and SciTech Premium Collection (ProQuest).