Experiments towards finding conditions for identification of metalated peptide sequences derived from transport proteins by MALDI TDS (top-down sequencing) method

Available Positions: 1

Evaluation Criteria:

• 1st Priority: Students of 1st Cicle in Biochemistry who have never attended the CQM internships;
• 2nd Priority: Students of the 2nd Cycles in Applied Biochemistry or Nanochemistry and Nanomaterials who have never attended the CQM internships;
• 3rd Priority: Students from other universities attending Biochemistry, Chemistry or related courses;
• 4th Priority: Other students of the 1st Cycle of Biochemistry and the 2nd Cycles in Applied Biochemistry or Nanochemistry and Nanomaterials;
• Candidates not selected in the first or second choices will be considered substitutes.

Application Deadline: 2019-06-16 17:00:00 (GTM/UTC + 0)

Reference: CQM-19-SUM-INT-01

Registration from is available here until 16^th June 2019. Applicants must indicate the preferred training courses taking into account that it is not possible to repeat the training course or supervisor from the previous year(s). Consider the observation(s) and other requirements indicated for each proposal.

Position: Experiments towards finding conditions for identification of metalated peptide sequences derived from transport proteins by MALDI TDS (top-down sequencing) method

Reference: CQM-19-SUM-INT-01

Position Type: Internship

Place of Work: CQM (Centro de Química da Madeira)

Host Institution: CQM (Centro de Química da Madeira)

Scientific Areas: Analytical Chemistry, Chemistry, Biochemistry.

Supervisor(s): Marijana Petkovic .

Experimental part of the work will be divided into following sections:

• incubation of selected transition metal complex with bovine serum albumin at various concentrations
• detection of incubation products by MALDI TOF TOF MS
• finding conditions for best detection of products
• in source decay fragmentation of BSA and metallated BSA by MALDI TOF TOF device after application of a compound at various matrices and at varying sample/matrix ratios.

It is expected for the student to develop the feeling about the scientific work in the field of biochemistry/mass spectrometry and to identify at least one target sequence at BSA for transition metal complexes.

Transition complexes (metallodrugs), such as cisplatin, are well known anti-tumor agents, which are successfully used over decades for therapy of various cancers. Being aggressive and less selective, most metallo-drugs induce severe side effects in patients, therefore, scientists make an effort to develop new generations of metallo-drugs which will kill cancer cells much more efficiently and with less side effects.

After administration to patient's circulation, a metallodrug interacts with serum transport proteins, and flows towards a diseased tissue, where it has been taken up by cancer cells using overexpressed receptors for transferrin and triggering endocytosis, or by passively through the plasma membrane. After the uptake, metallodrug activates and interacts with cellular target molecules, as well as with intracellular pathways leading to cell cycle arrest and/or inducing cellular apoptosis. However, the nature of interaction between the transport protein and metallodrug, is crucial, since it determines the concentration of a drug in the target tissue. Because the metallodrug binds after hydrolysis to the protein via electrostatic interactions with negatively charged amino acid residues, metallated protein and peptides might not be easily detectable by mass spectrometry, as many ionization approaches lead to the bond destruction. Only very recently, by using modifiers, the protein target sequences contained in serum albumin and transferrin to which certain ruthenium complexes bind could be discovered by mass spectrometric approach (mostly with electrospray ionization).

Matrix-assisted laser desorption and ionization time of flight mass spectrometry (MALDI TOF MS) is a soft ionization technique, which is suitable for analysis of proteins and peptides. Newer devices enable also top-down-sequencing of proteins, the approach which uses higher laser energies to induce fragmentation of a protein. This method was not routinely used for identification of target sequences on proteins, and therefore, the approach has to be first developed and then applied to selected proteins.

These training courses are free for the students of 1^st Cycle of Biochemistry and 2^nd Cycles of Applied Biochemistry or Nanochemistry and Nanomaterials of the University of Madeira.

For students of Biochemistry, Chemistry or similar areas from other universities the registration costs 50€. Selected candidates must have an insurance for individual accidents in order to use the laboratory areas of CQM.

The trainees will receive a participation certificate which will include the total worked hours.

NOTE: These internships do not replace any curricular equivalency / frequency of their courses.

Application Submission: 2019-06-06 09:00:00 until 2019-06-16 17:00:00 (GTM/UTC + 0)

Publication of Results: 2019-06-17

Admittance lists will be announced from 17th June 2019.

Contract Duration: 2019-07-02 to 2019-07-16

Daily Work Hours: 07:00:00

• 1st Priority: Students of 1st Cicle in Biochemistry who have never attended the CQM internships;
• 2nd Priority: Students of the 2nd Cycles in Applied Biochemistry or Nanochemistry and Nanomaterials who have never attended the CQM internships;
• 3rd Priority: Students from other universities attending Biochemistry, Chemistry or related courses;
• 4th Priority: Other students of the 1st Cycle of Biochemistry and the 2nd Cycles in Applied Biochemistry or Nanochemistry and Nanomaterials;
• Candidates not selected in the first or second choices will be considered substitutes.

Registration from is available here until 16^th June 2019. Applicants must indicate the preferred training courses taking into account that it is not possible to repeat the training course or supervisor from the previous year(s). Consider the observation(s) and other requirements indicated for each proposal.