Biochemistry 3381A Chapter Notes - Chapter 5.3, 5.5: Polyethylene Glycol, Methionine, Lactone
9 Oct 2017
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Protein techniques, cysteine chemistry, protein sequence/mass determination, fragmentation, electrophoresis (parts of sects. In vacuum, f would cause molecule to accelerate: f = eq/d -> e = voltage (electric potential), d = distance between electrodes. In solution, molecule experiences frictional drag ff r = radius of charged velocity molecule, n = viscosity of solution, v : molecular velocity proportional to charge and voltage. Inversely proportional to viscosity of medium and distance between electrodes: carried out in porous matrix (polyacrylamide/agarose) which slows movement according to molecule size. Two-dimensional gel electrophoresis: use isoelectric focusing and sds-page: protein separated on a gel according to pi, then electrophoresed in sds polyacrylamide gel, separating according to size, stain gel to visualize location of individual proteins. Hydrophobic interaction chromatography: proteins pass through column with hydrophobic groups covalently linked to it, e. g. phenyl sepharose is a matrix with attached phenyl groups.
