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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

 

 

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 Nr.:1.1.1.2/VIAA/1/16/144

Period: 36month (1st September 2017–31st August 2020)

Projectcosts: 133 806,00 EUR

Projectimplementer: 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.




Mājas lapas izstrādi finansēja ERAF 2.1.1.2. aktivitātes projekts Nr. 2010/0196/2DP/2.1.1.2.0/10/APIA/VIAA/004 "Latvijas biomedicīnas pētījumu integrācija Eiropas zinātnes telpā".