This
course is for scientists who are not familiar with techniques
of protein isolation and characterization. It is a rigorous
program that includes laboratory work all day and a lecture
with discussion session every evening. Each student will become
familiar with each of the major techniques in protein purification
by actually performing four separate isolations including:
(i) a regulatory protein from muscle tissue: (ii) a sequence-specific,
DNA binding protein: (iii) a recombinant protein overexpressed
in E. coli; and (iv) a membrane-bound receptor. A variety
of bulk fractionation, electrophoretic, and chromatographic
techniques will include: precipitation by salts, pH, and ionic
polymers; ion exchange, gel filtration, hydrophobic interaction,
and reverse phase chromatography; lectin affinity, ligand
affinity, oligonucleotide affinity, and immunoaffinity chromatography;
polyacrylamide gel electrophoresis, and electroblotting; and
high performance liquid chromatography. Procedures will be
present for solubilizing proteins from inclusion bodies and
refolding them into active monomeric forms. Methods of protein
characterization will be utilized to include immunological
and biochemical assays, peptide mapping, amino acid analysis,
protein sequencing, and mass spectrometry. Emphasis will be
placed on strategies of protein purification and characterization.
Guest lecturers will discuss protein structure, modification
of proteins, methodologies for protein purification and characterization,
and applications of protein biochemistry to cell and molecular
biology.
Speakers in the 2004 course
included:
Gerald Hart, Johns Hopkins University School of Medicine
Leemor Joshua-Tor, Cold Spring Harbor Laboratory
Sheenah Mische, Boehringer Ingelheim Pharmaceuticals, Inc.
Tom Muir, Rockefeller University
Bruce Stillman, Cold Spring Harbor Laboratory
This
course is supported with funds provided by the National
Cancer Institute
Cost
(including board and lodging): $2,665
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