S5017 Mass Spectrometry in Proteomics

Přírodovědecká fakulta
jaro 2025
Rozsah
1/0/0. Two-hour blocks. 1 kr. (plus ukončení). Doporučované ukončení: k. Jiná možná ukončení: zk.
Vyučováno kontaktně
Vyučující
Mgr. Gabriela Lochmanová, Ph.D. (přednášející)
prof. RNDr. Zbyněk Zdráhal, Dr. (přednášející)
Garance
prof. RNDr. Zbyněk Zdráhal, Dr.
Národní centrum pro výzkum biomolekul – Přírodovědecká fakulta
Kontaktní osoba: RNDr. Michaela Fajkusová
Dodavatelské pracoviště: Středoevropský technologický institut
Předpoklady
Basic knowledge of biochemistry and molecular biology
Omezení zápisu do předmětu
Předmět je otevřen studentům libovolného oboru.
Cíle předmětu
The aim of this course is to introduce to students present possibilities of mass spectrometry in proteomics, the basic principles of mass spectrometry, types of MS instrumentation, basics of interpretation of MS and MS/MS data and using examples to present application of appropriate mass spectometric techniques for different types of biological and biomedical experiments.
Výstupy z učení
At the end of the course students should be able to: understand and explain the basic principles of mass spectrometry; describe MS instrumentation currently used in proteomics; understand basics of interpretation of MS and MS/MS data; explain principles of protein identification and their posttranslational modifications by mass spektrometry; explain principles of MS quantification methods; describe present possibilities of mass spectrometry in proteomics, propose application of appropriate mass spectrometric techniques for different types of biological and biomedical experiments.
Osnova
  • 1. Mass spectrometry (MS) Method principles, method possibilities in proteomics, overview of MS instrumentation and ionization techniques (MALDI-MS, ESI-IT MS, FTMS, hybrid systems).
  • 2. Sample preparation and separation (RNDr. Hana Konečná) General rules for sample preparation before MS analysis. Basic techniques for protein isolation. Overview of separation techniques (electromigration and chromatographic techniques).
  • 3. Sample preparation and separation II (Dr. Gabriela Lochmanová) Strategies for processing of protein samples (selection of appropriate techniques in consideration of purpose of experiment).
  • 4. Basic methods of protein identification Proteomic approaches (bottom-up, top-down,)peptide mapping, MS/MS ion search, identification of proteins with unknown sequence(de-novo sequencing), protein homology.
  • 5. Protein quantification Methods of relative and absolute quantification in mass spectrometry (mass labels, isobaric labels-iTRAQ, TMT, label-free approach).
  • 6. Characterization of protein modifications Overview of modification types (chemical, posttranslational modifications), mutations. Phosphoproteins - sample preparation and detection, localization of phosphorylations. Glycoproteins sample preparation and detection, glycan analysis.
  • 7. Proteomic applications Mass spectrometry and disease diagnostics (biomarker identification, protein profiling), proteomic projects in our proteomic facility.
Literatura
  • Proteome research : mass spectrometry. Edited by Peter James. Berlin: Springer-Verlag, 2001, xxi, 274 s. ISBN 3-540-67255-9-. info
Výukové metody
lecture, class discussion
Metody hodnocení
colloquium or oral examination
colloquium: three questions for each student, correct answers to all questions are needed to pass
oral examination: questions and discussion, correct answers to questions (at least 60%) and understanding discussed topics is needed to pass the exam
Vyučovací jazyk
Angličtina
Další komentáře
Předmět je vyučován každoročně.
Výuka probíhá blokově.
Poznámka k četnosti výuky: Two-hour blocks.
Předmět je zařazen také v obdobích jaro 2022, jaro 2023, jaro 2024.