Journal Article

Production, purification, and quality assessment of borrelial proteins CspZ from Borrelia burgdorferi and FhbA from Borrelia hermsii

Avalle, Bérangère, Marquant, Rodrigue, Guérin, Mickaël, Padiolleau-Lefèvre, Séverine, Prost, Elise, Vandevenne, Marylène, Octave, Stéphane, Brans, Alain, Matagne, André, Maffucci, Irene

2024

Overview

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 FH 15-20  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 Graphical Abstract

• Publisher: Springer Berlin Heidelberg,Springer,Springer Nature B.V,Springer Verlag,Springer Science and Business Media Deutschland GmbH
• Subject: Affinity / Affinity chromatography / Alternative pathway / Applied Microbiology and Biotechnology / Bacterial Proteins / Bacterial Proteins - chemistry / Bacterial Proteins - genetics / Bacterial Proteins - metabolism / Bacteriology / bioactive properties / Biochemistry, biophysics & molecular biology / Biochemistry, Molecular Biology / Biochimie, biophysique & biologie moléculaire / Biological activity / Biomedical and Life Sciences / Biotechnologically Relevant Enzymes and Proteins / Biotechnology / blood serum / Borrelia / Borrelia - genetics / Borrelia - immunology / Borrelia - metabolism / Borrelia burgdorferi / Borrelia burgdorferi - genetics / Borrelia burgdorferi - immunology / Borrelia burgdorferi - metabolism / Borrelia hermsii / Borreliosis / Chromatography, Affinity / Circular dichroism / circular dichroism spectroscopy / Complement / Complement C3b Inactivator Proteins / Complement C3b Inactivator Proteins - genetics / Complement C3b Inactivator Proteins - metabolism / Complement Factor H / Complement Factor H - genetics / Complement Factor H - metabolism / Complement system / CspZ / Diagnosis / Dichroism / E coli / Escherichia coli / Escherichia coli - genetics / Escherichia coli - metabolism / etiology / Factor H / Factor H binding A / Factor-H like 1 / fever / FhbA / FHL-1 protein / Gene Expression / Genomics / H-binding / High-performance liquid chromatography / Humans / Immune evasion / Jarisch-Herxheimer reaction / Life Sciences / Liquid chromatography / Lyme Disease / Lyme Disease - microbiology / Mass spectrometry / Mass spectroscopy / Microbial Genetics and Genomics / Microbiology / Microbiology and Parasitology / NMR / Nuclear magnetic resonance / Physiological aspects / polyacrylamide gel electrophoresis / Protein folding / Protein purification / Protein quality control / protein value / Proteins / Quality assessment / Quality control / Recombinant Proteins / Recombinant Proteins - chemistry / Recombinant Proteins - genetics / Recombinant Proteins - isolation & purification / Recombinant Proteins - metabolism / Relapsing fever / Sciences du vivant / Spirochetes / Surface plasmon resonance / Surface proteins / Tick-borne diseases / Ticks / Vertebrates