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Struktura, biochemia, fizjologia i ewolucja bakteryjnych α2-makroglobulin z ludzkich periodontopatogenów Porphyromonas gingivalis i Tannerella forsythia (Supervisor: Mirosław Książek)

Struktura, biochemia, fizjologia i ewolucja bakteryjnych α2-makroglobulin z ludzkich periodontopatogenów Porphyromonas gingivalis i Tannerella forsythia (Supervisor: Mirosław Książek)

Admission information:

Pursuant to § 7 sections 1 and 3 of Resolution No. 91/IX/2021 of the Senate of the Jagiellonian University of 29 September 2021 on: the rules of recruitment for the Doctoral School of Exact and Natural Sciences at the Jagiellonian University in the academic year 2022/2023, the Director of the Doctoral School of Exact and Natural Sciences announces a competition for one doctoral student/s with a scholarship financed
from the research project funds as part of the PhD study programme Biochemistry, biophysics, molecular biology and biotechnology.
Within the project Sonata Bis 11, financed by the National Science Center entitled “The structures, biochemistry, physiology, and evolution of bacterial α2-macroglobulins from the human periodontopathogens Porphyromonas gingivalis and Tannerella forsythia”, a commission set up at the School will conduct recruitment for the PhD study programme Biochemistry, biophysics, molecular biology and biotechnology.
The project manager, Mirosław Książek, PhD and Department of Microbiology offers an opportunity to complete the doctorates on the biochemical, structural and functional characterisation of bacterial α2-macroglobulins derived from human pathogens. In the project will be used techniques in the field of biochemistry, structural biology and microbiology, such as cryogenic electron microscopy, X-ray crystallography, flow cytometry or kinetic characterisation of inhibition of proteases by inhibitors.
After the interviews and the evaluation of the candidates, the Chairman of the committee presents a report to the Director containing a list of candidates recommended for admission to the school as part of the research project.

Project description:

The main aim of the proposed project is not only a detailed biochemical and structural description of the inhibitory mechanism of the α2-macroglobulin (A2M) produced by two human periodontal pathogens: Porphyromonas gingivalis, PgA2M and Tannerella forsythia, TfA2M, but also to describe the role of these A2Ms in virulence of these bacteria.
TfA2M and PgA2M will be obtained as recombinant proteins employing a bacterial expression system during the project. First, we will identify human and endogenous proteases inhibited by investigated A2Ms and biochemically characterise the inhibition process in detail. To achieve it, we will determine the stoichiometry of inhibition (SI), the number of A2M molecules required for inhibition of one molecule of active protease, and association rate constant (kass), a parameter describing how fast protease activity is scavenged, identify the molecular mass of smallest proteinaceous substrate for which inhibition occurred, and analyse the formation and chemical nature of inhibitory complexes. In this step, we will also analyse the inhibitory flexibility of A2Ms by introducing different changes to A2M, aiming to improve the inhibitory properties of A2Ms. Secondly, we will solve the three-dimensional structure of A2Ms: native and in complex with target proteases employing X-ray crystallography and cryogenic electron microscopy (cryo-EM). Finally, we will describe the biological role of A2Ms. To do it, we will check using P. gingivalis and T. forsythia cells: wild-type and deletion mutants lacking A2M if A2Ms protect the bacteria against killing by human phagocytic (neutrophils, macrophages) and epithelial cells. We will also check if A2Ms, through inhibition of a protease, dentilisin, limit virulence of another periodontopathogen, Treponema denticola.
 

Candidate’s profile:

  • experience in molecular biology (e.g. PCR, molecular cloning, mutagenesis) and working with bacterial cells (e.g., growing them, ideally, but not necessary, production of recombinant proteins), or growing and performing experiments with eukaryotic cells; 
  • practical or at least academic (from the practical courses during the studies) knowledge of basic protein techniques (e.g., SDS-PAGE, measurement of enzymatic activity, western blot); 
  • good knowledge of English.

Schedule of the competition:

  1. Opening of the competition (means: opening admission in the Online Application System): 27.05.2022
  2. Application submission deadline: (means: closing the Online Application System) 22.06.2022
  3. Entrance exams: 29.06-04.07.2022
  4. Announcement of results: 11.07.2022
  5. Enrolment: 12.07-15.09.2022
  6. Enrollment (reserve list): 16-27.09.2022
 

Application documents:

curriculum vitae (CV).
 
Please send your application documents to the project manager Mirosław Książek, PhD at (e-mail) miroslaw.ksiazek@uj.edu.pl and upload them into the Online Application System (irk.uj.edu.pl).

Online Application link

Documents required for entry within PhD study programme Biochemistry, biophysics, molecular biology and biotechnology:
  • the original OAS application printed out from the system, containing a photograph;
  • a photocopy and the original for inspection of: a diploma or other document of graduation of a higher education institution abroad entitling the holder to undertake education in order to obtain the degree of doctor in which it was issued, recognised as equivalent to the relevant Polish master's diploma, and if, by the date of entry, the candidate does not yet hold a master's diploma document, a certificate issued by the higher education institution confirming that the candidate has passed the diploma examination, containing information on the right to take up education in order to obtain the degree of doctor in whose system the institution operates, with the grades given from the diploma examination, from the diploma thesis and on the diploma (for graduates of the second degree studies also a certificate containing these grades from the completed first degree studies);
  • a photocopy and the original for inspection: a diploma supplement;
  • for inspection: identity document (passport in case of foreigners);
  • originals of all other documents provided at the registration stage in the form of a scan.
In the event of exposure to harmful, noxious or hazardous factors, the applicant will, at the time of registration, be referred for a medical examination by an occupational medicine physician. The candidate shall be obliged to deliver, within the time limit set by the programme manager, a medical certificate confirming the absence of contraindications to take up education (applies only to some PhD study programmes).
 
Higher education diplomas awarded abroad should:
 
  1. bear an apostille clause if the issuing country is covered by the Convention abolishing the requirement legalisation for foreign public documents, done at The Hague on 5 October 1961 (Journal of Laws of 2005, No. 112, item 938) or
  2. be subjected to legalisation, in other cases.
Any document submitted in a language other than Polish or English must be accompanied by a certified translation into Polish or English.
 

Insurance:

Each doctoral student, including a foreigner, admitted to the doctoral school is obligatorily subject to health insurance if he/she is not covered by such insurance on other grounds (e.g. employment relationship, contract of mandate, business activity, insured as a family member under 26 years of age, as the spouse of an insured person). The health insurance contribution is paid by the Jagiellonian University and is financed from the State budget. In addition, a doctoral student receiving a doctoral scholarship is subject to compulsory retirement and disability insurance and accident insurance.
 

IMPORTANT:

A person admitted to the doctoral school begin education and acquire doctoral student rights upon taking the oath. The Department of Personal Affairs reports each doctoral student admitted to the School to compulsory health and social insurance at the commencement of education. Until that time foreign candidates are obliged to cover the costs of insurance for the time of travel, treatment, etc. individually.
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