[en] Borrelia, spirochetes transmitted by ticks, are the etiological agents of numerous multisystemic diseases, such as Lyme borreliosis (LB) and tick-borne relapsing fever (TBRF). This study focuses on two surface proteins from two Borrelia subspecies involved in these diseases: CspZ, expressed by Borrelia burgdorferi sensu stricto (also named BbCRASP-2 for complement regulator-acquiring surface protein 2), and the factor H binding A (FhbA), expressed by Borrelia hermsii. Numerous subspecies of Borrelia, including these latter, are able to evade the immune defenses of a variety of potential vertebrate hosts in a number of ways. In this context, previous data suggested that both surface proteins play a role in the immune evasion of both Borrelia subspecies by interacting with key regulators of the alternative pathway of the human complement system, factor H (FH) and FH-like protein 1 (FHL-1). The recombinant proteins, CspZ and FhbA, were expressed in Escherichia coli and purified by one-step metal-affinity chromatography, with yields of 15 and 20 mg or pure protein for 1 L of cultured bacteria, respectively. The purity was evaluated by SDS-PAGE and HPLC and is close to about 95%. The mass of CspZ and FhbA was checked by mass spectrometry (MS). Proper folding of CspZ and FhbA was confirmed by circular dichroism (CD), and their biological activity, namely their interaction with purified FH from human serum (recombinant FH15-20 and recombinant FHL-1), was characterized by SPR. Such a study provides the basis for the biochemical characterization of the studied proteins and their biomolecular interactions which is a necessary prerequisite for the development of new approaches to improve the current diagnosis of LB and TBRF. KEY POINTS: • DLS, CD, SEC-MALS, NMR, HPLC, and MS are tools for protein quality assessment • Borrelia spp. possesses immune evasion mechanisms, including human host complement • CspZ and FhbA interact with high affinity (pM to nM) to human FH and rFHL-1.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Guérin, Mickaël; Unité de Génie Enzymatique et Cellulaire (GEC), CNRS UMR 7025, Université de Technologie de Compiègne, Compiègne, 60203, France
Brans, Alain ; Université de Liège - ULiège > Département des sciences de la vie > Centre d'Ingénierie des Protéines (CIP)
Matagne, André ; Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines
Marquant, Rodrigue; Unité de Génie Enzymatique et Cellulaire (GEC), CNRS UMR 7025, Université de Technologie de Compiègne, Compiègne, 60203, France
Prost, Elise; Unité de Génie Enzymatique et Cellulaire (GEC), CNRS UMR 7025, Université de Technologie de Compiègne, Compiègne, 60203, France
Octave, Stéphane; Unité de Génie Enzymatique et Cellulaire (GEC), CNRS UMR 7025, Université de Technologie de Compiègne, Compiègne, 60203, France
Avalle, Bérangère; Unité de Génie Enzymatique et Cellulaire (GEC), CNRS UMR 7025, Université de Technologie de Compiègne, Compiègne, 60203, France
Maffucci, Irene; Unité de Génie Enzymatique et Cellulaire (GEC), CNRS UMR 7025, Université de Technologie de Compiègne, Compiègne, 60203, France
Padiolleau-Lefèvre, Séverine ; Unité de Génie Enzymatique et Cellulaire (GEC), CNRS UMR 7025, Université de Technologie de Compiègne, Compiègne, 60203, France. severine.padiolleau@utc.fr
Language :
English
Title :
Production, purification, and quality assessment of borrelial proteins CspZ from Borrelia burgdorferi and FhbA from Borrelia hermsii.
Publication date :
23 July 2024
Journal title :
Applied Microbiology and Biotechnology
ISSN :
0175-7598
eISSN :
1432-0614
Publisher :
Springer Science and Business Media Deutschland GmbH, Germany
MESRI - Ministère de l’Enseignement supérieur, de la Recherche et de l’Innovation Sorbonne Université
Funding text :
This study was supported by the French Ministry of Higher Education, Research and Innovation (MESRI), Sorbonne University Idex as part of the French government\u2019s support for the \u201CInvestissements d\u2019Avenir\u201D programms, Lyme Support endowments funds, and by the Institut des Sciences du Calcul et des Donn\u00E9es (ISCD) of Sorbonne University (IDEX SUPER 11-IDEX-0004). Equipment is supported by the Regional Council of Picardie and European Union (grant number CPER 2007\u20132020). The funding bodies played no role in the design of the study, collection, analysis, interpretation of data, nor in the writing of the manuscript.
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