C7250 Protein characterisation using mass spectrometry

Faculty of Science
Autumn 2014
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)
RNDr. Hana Konečná (lecturer)
prof. RNDr. Šárka Pospíšilová, Ph.D. (lecturer)
Mgr. Gabriela Lochmanová, Ph.D. (lecturer)
Guaranteed by
prof. RNDr. Jiří Fajkus, CSc.
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
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 spectrometry explain principles of MS quantification methods describe present possibilities of mass spectrometry in proteomics propose application of appropriate mass spectometric techniques for different types of biological and biomedical experiments
Syllabus
  • 1. Mass spectrometry (MS)
  • Method principles, method possibilities, 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. Strategies for processing of protein samples (selection of appropriate techniques in consideration of purpose of experiment).
  • 3. Basic methods of protein identification
  • Peptide mapping, MS/MS ion search, identification of proteins with unknown sequence(de-novo sequencing), basic proteomic approaches (bottom-up, top-down), protein homology.
  • 4. Quantification
  • Methods of relative and absolute quantification in mass spectrometry (mass labels, isobaric labels-iTRAQ).
  • 5. Determination of protein modifications
  • Overview of modification types (chemical, post-translational modifications), mutations. Phosphoproteins - sample preparation and detection, localization of phosphorylations. Glycoproteins sample preparation and detection, glycan analysis.
  • 6. Protein arrays (RNDr. Šárka Pospíšilová, PhD)
  • Protein array types, their preparation, detection a quantification.
  • 7. Proteomic applications
  • Mass spectrometry and disease diagnostics (biomarker identification, protein profiling), proteomic projects in our Core lab.
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 2015, Autumn 2016, autumn 2017, Autumn 2018, Autumn 2019, Autumn 2020, autumn 2021, Autumn 2022, Autumn 2023, Autumn 2024.
  • Enrolment Statistics (Autumn 2014, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2014/C7250