Proteomics course: gel based protein separation by twodimensional
gel
electrophoresis
and
protein
characterization by MALDI-TOF/TOF mass spectrometry
18 - 21 June 2012
CIIMAR - Centre of Marine and Environmental Research, University of Porto
IPATIMUP - Institute of Molecular Pathology and Immunology of the University of Porto
FCUP - Faculty of Sciences, University of Porto
Introduction: Proteomics is a research area devoted to the analysis of proteins from
complex biological samples. It has been applied to evaluate protein expression, to identify
proteins and characterize their post-translational modifications (PTMs). Examples of such
applications include the analysis of the biochemical changes in organisms when exposed
to an adverse environment, allowing therefore a better understanding on organism’s
response and adaptation. Two main approaches to undertake proteomic studies are the
two-dimensional gel electrophoresis of proteins (2DE) and MALDI-TOF/TOF mass
spectrometry. The development of the two methodologies has allowed high throughput
proteomic analyses and PTM research.
Course aims: This 4 day course aims to describe the principles of 2DE and MALDI-TOF/TOF
as well as to provide basic training in the two methodologies. The course will be held at
CIIMAR and IPATIMUP, being organized by the Laboratory of Ecotoxicology, Genomics and
Evolution (LEGE/CIIMAR) and the IPATIMUP Proteomics Unit. It will consist in theoretical
lectures and laboratory work on protein separation by 2DE, peptide analyses, protein
identification and characterization of post-translational modifications by MALDI-TOF/TOF
mass spectrometry.
Participants: graduated students and researchers interested in proteomics research
and/or to gain practical skills in 2DE and MALDI-TOF/TOF mass spectrometry analyses. The
course will be limited to 12 Participants. For more information contact Alexandre Campos,
acampos@ciimar.up.pt.
Organization:
Alexandre Campos (Laboratory of Ecotoxicology, Genomics and Evolution, CIIMAR, FCUP,
University of Porto).
Hugo Osório (Proteomics Unit, IPATIMUP).
Vitor Vasconcelos (Faculty of Sciences, University of Porto and CIIMAR).
André Almeida (Instituto de Investigação Científica Tropical, IICT).
Pedro Rodrigues (University of Algarve)
Preliminary Program:
18 June:
9.00 am – 13.00 pm: lectures (CIIMAR)
9.00 Welcome reception
9.30 Introduction to proteomics A. Almeida
10.10 Principles of protein sample preparation and 2DE A. Campos
10.50 Coffee break
11.10 Principles of MALDI-TOF/TOF MS and protein identification H Osorio
11.50 Protein quantification in proteomics
12.30 General discussion
13.00 Lunch
14.30 pm – 18.00 pm: laboratory work (CIIMAR)
Sample preparation and protein separation by Isoelectric Focusing (IEF) (First dimension)
19 June:
8.30 am – 13.00 pm: laboratory work (IPATIMUP)
- Protein isolation and “in gel” digestion
- Sample application in the MALDI plate
- Mass spectra acquisition in MS and MS/MS modes
- Mass spectra interpretation and protein identification
14.30 pm – 18.00 pm: lectures (CIIMAR)
Four lectures given by invited researchers. Thirty minutes presentations and discussion.
Coffee break (30 minutes):
David Eckersall - 'Proteomics in Animal Health and Disease'
Ingrid Miller - How to arrive at good quality gels and gel data
Susana Cristobal (to be announced)
Pedro Rodrigues - Assessing fish muscle quality using proteomics. Validation of a sample
fractionation procedure
20 June:
8.30 am – 13.00 pm: laboratory work (CIIMAR/IPATIMUP)
- Second dimension SDS-PAGE
- Protein identification by homology search
14.30 pm – 18.00 pm: lectures (CIIMAR)
Four lectures given by invited researchers. 30 minutes presentations followed by
discussion. Coffee break of 30 minutes.
Flemming Jessen - 2DE based proteomics for prediction and understanding of
seafood quality
Jarlath
Nally
-
Gel
electrophoresis
and
immunoblotting
of
bacterial
membrane proteins
Kristin Hollung - Proteomics as a research tool in meat science
Miguel A. Sentandreu – The use of Peptide biomarkers in meat composition analysis
21 June:
8.30 am – 13.00 pm: laboratory work (CIIMAR)
Demonstrations of gel image analysis and relative quantification of proteins using
SameSpots software (NonLinear Dynamics).