Funding: European Regional Development Fund (ERDF) “On Implementation of Activity 1.1.1.2 “Post-doctoral Research Aid” of the Specific Aid Objective 1.1.1 “To increase the research and innovative capacity of scientific institutions of Latvia and the ability to attract external financing, investing in human resources and infrastructure” of the Operational Programme “Growth and Employment”
Project Title: „Structural and functional studies of Lyme disease agent Borrelia burgdorferi outer surface proteins to reveal the mechanisms of pathogenesis with the intention to create a new vaccine”
Project No.: 1.1.1.2/VIAA/1/16/144
Period: 36month (1st September 2017–31st August 2020)
Project costs: 133 806,00 EUR
Project implementer: Dr.biol. K. Brangulis
The goal of this Project is to solve the 3-d structures of Lyme disease agent B. burgdorferi outer surface proteins predicted to be important in the pathogenesis of Lyme disease to reveal the molecular details of action that could be used in future to design new strategies to stop the infection.
Objectives and overview of the action – to select the target B. burgdorferi outer surface proteins and to make expression vector constructs. The selection criteria for the target proteins will be based on the importance in the pathogenesis of Lyme disease as judged from the scientific articles.
However, it must be emphasized that for most of the up-regulated B. burgdorferi outer surface proteins known to be potentially important in the pathogenesis of Lyme disease, the exact function/ligand/receptor is unknown, and they do not possess any homology with proteins in any other organism whose genome has been sequenced, that also prevents to make any assumption about the potential function. Therefore, a comprehensive study of outer surface proteins is required to reveal the molecular details of pathogenesis of Lyme disease which remains poorly understood. It will certainly provide a significant contribution in the understanding of the molecular details of Lyme disease action but most importantly is that the acquired knowledge about the molecular details of outer surface proteins is intended to be used to develop new strategies to fight against the Lyme disease in humans.
Information published: 01.09.2017.
Progress of the project
1 September 2017 – 30 November 2017
In the current period the project was carried out according to the project time diagramma, i.e., the research has been dedicated to perform according to work packages 1 and 2 (WP1 and WP2). WP1 and WP2 include the selection of target Borrelia burgdorferi outer surface proteins for design of expression constructs and production of proteins in Escherichia coli cells and purification of expressed borrelial proteins.
In the period until 30.11.2017 15 different B. burgdorferi outer surface proteins which have showed some role in the pathogenesis of Lyme disease have been selected. 15 different expression vector constructs for protein expression in E. coli have been made accordingly. 5 different proteins have been expressed in high quantities, cells have been sonicated and all 5 proteins have been purified using chromatography methods. Crystallization trials have been started for 2 proteins.
Information published: 30.11.2017.
Progress of the project
1 December 2017 – 28 February 2018
In the current period from 01.12.2017 to 28.02.2018 the project was carried out according to the project time diagramma. The research was conducted according to the work packages 1-4, which consists of research activities to select the target B. burgdorferi proteins, to make expression constructs, to express the proteins in Escherichia coli, to purify the target proteins by chromatography methods, to crystallize the protein, to collect the diffraction data and to solve the 3-d structure.
In the period from 01.12.2017 to 28.02.2018 6 different B. burgdorferi proteins were successfully expressed in E. coli, and purified by affinity and ion-exchange chromatography methods. 5 different proteins were crystallized and the promising conditions were optimized. Diffraction data were collected for 2 different B. burgdorferi proteins and the 3-d structure was determined.
Information published: 28.02.2018.
Progress of the project
1 March 2018 – 31 May 2018
In the current period from 01.03.2018 to 31.05.2018 the project was carried out according to the project time diagramma. 5 different B. burgdorferi outer surface proteins have been expressed in E. coli. Afterwards the proteins have been purified by using chromatography methods (affinity and ion-exchange), crystallization screenings have been set and the conditions have been optimized. Diffraction data from crystals for 3 different B. burgdorferi proteins has been collected at the synchrotron, including Se-Met derived protein crystals. The work on diffraction data processing and structure determination is in progress.
Information published: 31.05.2018.
Progress of the project
1 June 2018 – 31 August 2018
The project was carried out according to the project time diagram. In the current period several B. burgdorferi outer surface proteins were purified by chromatography, crystallized and the favourable crystallization conditions optimized. By using the previously collected diffraction data the crystal structures for two different proteins were determined by molecular replacement and SAD technique. A scientific article has been published and work on a new publication has started.
Information published: 31.08.2018.
Progress of the project
1 September 2018 – 30 November 2018
The project was carried out according to the project time diagramma. In the current period 6 different B. burgdorferi Se-Met derived proteins were obtained. The proteins were successfully purified by chromatography methods. The purified proteins were crystallized and the positive conditions were optimized. Crystals suitable for diffraction data collection were flash-frozen in liquid nitrogen. From previously collected diffraction data, a new 3-d structure has been solved.
Information published: 30.11.2018.
Progress of the project
1 December 2018 – 28 February 2019
The project was carried out according to the project time diagramma. Several B. burgdorferi proteins were crystallized and the crystallization conditions optimized. At the BESSY II synchrotron the diffraction data were collected and the 3-d structures were solved. Analysis of the obtained structural data was carried out and a scientific article is in preparation.
Information published: 28.02.2019.
Progress of the project
1 March 2019 – 31 May 2019
The work has been carried out on solving the 3-D structures of Lyme disease causing agent Borrelia burgdorferi by using the diffraction data collected at the BESSY II synchrotron (Germany). An article has been written about the 3-D structure of B. burgdorferi protein BB0365 that was solved during the current project. Several B. burgdorferi Se-Met derived proteins have been crystallized and the crystals have been collected and stored in liquid nitrogen.
Information published 31.05.2019.
Progress of the project
1 June 2019 – 31 August 2019
In the current period crystallization conditions were optimized for 4 different B. burgdorferi outer surface proteins. Interaction and functional studies of B. burgdorferi protein BBE31 with isothermal titration calorimetry were performed. Two scientific articles have been prepared for submission.
Information published 30.08.2019.
Progress of the project
1 September 2019 – 30 November 2019
Protein diffraction data analysis for previously obtained diffraction data was performed during the current project implementation period. Functional studies have been carried out on certain B. burgdorferi surface proteins. Structural analysis of the B. burgdorferi outer surface protein BBE31 has been published.
Information published 29.11.2019.
Progress of the project
1 December 2019 – 29 February 2020
During the respective project period, anomalous diffraction data for two different outer surface protein crystals of Borrelia burgdorferi were processed and also processing of native data for two different protein crystals was performed. Two scientific articles were submitted and published related to the protein 3D structures of Lyme disease agent that were obtained during the project – https://doi.org/10.1016/j.bbagen.2019.129499 and https://doi.org/10.1016/j.jsb.2020.107490.
Information published 28.02.2020.
Progress of the project
1 March 2020 – 31 Mat 2020
During the respective project implementation period, based on the solved crystal structures and structural similarities with homologous proteins, the work on functional studies of Borrelia burgdorferi outer surface proteins using chromatography and mass spectrometry methods has been carried out.
Information published 29.05.2020.
Progress of the project
1 June 2020 – 31 August 2020
During the respective project implementation period, work was performed on the processing of diffraction data in order to improve the quality of the respective 3-d structures and the data obtained during the project were summarized, which have resulted in a total of 6 publications – https://doi.org/10.1038/s41598-018-29651-9; https://doi.org/10.1016/j.ttbdis.2019.06.009; https://doi.org/10.1107/S2059798319010751; https://doi.org/10.1002/1873-3468.13594; https://doi.org/10.1016/j.bbagen.2019.129499; https://doi.org/10.1016/j.jsb.2020.107490.
Information published 31.08.2020.