Course: Practice in Proteomics

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Course title Practice in Proteomics
Course code LRR/CVPRO
Organizational form of instruction Exercise
Level of course Master
Year of study not specified
Semester Summer
Number of ECTS credits 3
Language of instruction Czech
Status of course Compulsory
Form of instruction Face-to-face
Work placements This is not an internship
Recommended optional programme components None
Lecturer(s)
  • Simerský Radim, Mgr. Ph.D.
  • Chamrád Ivo, Mgr. Ph.D.
  • Lenobel René, Mgr. Ph.D.
Course content
1. Extraction of a soluble fraction of proteins from a biological material; 1D or 2D SDS/PAGE gel electrophoresis; Commassie Briliant Blue gel staining. 2. Cutting of selected protein spots from a gel, in-gel digestion with trypsin and/or other proteases (e.g. chymotrypsin); extraction of digested peptides. 3. Preparation of peptide extracts for mass spectrometry analysis - Zip Tip purification on a C18 sorbent, a chemical modification of peptides with two different reagents; mass spectrometry analysis of purified and modified peptides and protein identification after database search (MALDI-TOF, ESI-QTOF, MASCOT DATABASE).

Learning activities and teaching methods
Laboratory Work
  • Homework for Teaching - 1 hour per semester
  • Attendace - 20 hours per semester
Learning outcomes
To learn practical methods used in proteomic.
Student should be able to (after attending the course): - apply basic proteomic methods for analysis of proteins in a biological material - analyse collected raw data (protein identification, determination of molecular mass, study of post-translational protein modification and others) - write an experimental protocol and discuss of obtained results
Prerequisites
Basic experiences with laboratory work, knowledge from proteomic lectures.

Assessment methods and criteria
Written exam

100% attendance, completion of all experimental tasks
Recommended literature
  • Keough T., Lacey M. P., Youngquist R. S. (2000). Derivatization procedures to facilitate de novo sequencing of lysine-terminated tryptic peptides using postsource decay matrix-assisted laser desorption/ionization mass spectrometry, Rapid Communication. Mass Spectrom. 14, 2348-2356.
  • Keough T., Youngquist R. S., Lacey M. P. (1999). A method for high-sensitivity peptide sequencing using postsource decay matrix-assisted laser desorption ionization mass spektrometry. Proc. Natl. Acad. Sci. USA 96, 7131-7136.
  • Marekov L. N., Steinert P. M. (2003). Charge derivatization by 4-sulfophenyl isothiocyanate enhances peptide sequencing by post-source decay matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. J. Mass Spectrom. 38, 373-377.
  • Pingoud A., Urbanke C., Hoggett J., Jeltsch A. (2002). Biochemical methods: A concise guide for students and researchers. Wiley-VCH Verlag GmbH, Weinheim, Germany.
  • Wang D., Kalb S. R., Cotter R. J. (2004). Improved procedures for N-terminal sulfonation of peptides for matrix-assisted laser desorption/ionization post-source decay peptide sequencing, Rapid Communication. Mass Spectrom. 18, 96-102.
  • Westermeier, R., Naven, T. (2002). Proteomics in practice. A laboratory manual of proteome analysis.. Wiley-VCH, Weinheim, Germany.
  • Winter M., Sherman N. E. (2000). Protein sequencing and identification using tandem mass spectrometry. John Wiley & Sons, Inc., New York, NY, USA.


Study plans that include the course
Faculty Study plan (Version) Category of Branch/Specialization Recommended year of study Recommended semester
Faculty: Faculty of Science Study plan (Version): Experimental Biology of Plants (2021) Category: Biology courses 1 Recommended year of study:1, Recommended semester: Summer
Faculty: Faculty of Science Study plan (Version): Experimental Biology (2015) Category: Biology courses 1 Recommended year of study:1, Recommended semester: Summer
Faculty: Faculty of Science Study plan (Version): Molecular and Cell Biology (2021) Category: Biology courses 1 Recommended year of study:1, Recommended semester: Summer