Production, purification, and quality assessment of borrelial proteins CspZ from Borrelia burgdorferi and FhbA from Borrelia hermsii
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 Borr...
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Published in | Applied microbiology and biotechnology Vol. 108; no. 1; p. 425 |
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Main Authors | , , , , , , , , , |
Format | Journal Article Web Resource |
Language | English |
Published |
Berlin/Heidelberg
Springer Berlin Heidelberg
01.12.2024
Springer Nature B.V Springer Science and Business Media Deutschland GmbH |
Subjects | |
Online Access | Get full text |
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Summary: | 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
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CspZ and FhbA interact with high affinity (pM to nM) to human FH and rFHL-1
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 scopus-id:2-s2.0-85199541262 |
ISSN: | 0175-7598 1432-0614 1432-0614 |
DOI: | 10.1007/s00253-024-13195-2 |