C7250 Protein characterisation using mass spectrometry

Faculty of Science
Autumn 2023
Extent and Intensity
1/0/0. 1 credit(s) (plus extra credits for completion). Recommended Type of Completion: k (colloquium). Other types of completion: zk (examination).
Teacher(s)
prof. RNDr. Zbyněk Zdráhal, Dr. (lecturer)
Mgr. Gabriela Lochmanová, Ph.D. (lecturer)
Guaranteed by
prof. RNDr. Zbyněk Zdráhal, Dr.
National Centre for Biomolecular Research – Faculty of Science
Contact Person: prof. RNDr. Zbyněk Zdráhal, Dr.
Supplier department: National Centre for Biomolecular Research – Faculty of Science
Timetable
Fri 8:00–9:50 C02/211
Prerequisites
Basic knowledge of biochemistry and molecular biology
Course Enrolment Limitations
The course is also offered to the students of the fields other than those the course is directly associated with.
fields of study / plans the course is directly associated with
Course objectives
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.
Learning outcomes
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.
Syllabus
  • 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.
Literature
  • Proteome research : mass spectrometry. Edited by Peter James. Berlin: Springer-Verlag, 2001, xxi, 274 s. ISBN 3-540-67255-9-. info
Teaching methods
lecture, class discussion
Assessment methods
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
Language of instruction
Czech
Further comments (probably available only in Czech)
Study Materials
The course is taught annually.
Listed among pre-requisites of other courses
The course is also listed under the following terms Autumn 2013, Autumn 2014, Autumn 2015, Autumn 2016, autumn 2017, Autumn 2018, Autumn 2019, Autumn 2020, autumn 2021, Autumn 2022, Autumn 2024.
  • Enrolment Statistics (Autumn 2023, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2023/C7250