BIOD60H3 Lecture Notes - Cyanogen Bromide, Carboxypeptidase B, Glutamine
25 Apr 2014
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Salting out: increase ionic strength increase solubility (salting in), then decrease (salting out: ionic charge: ion exchange chromatography electrophoresis isoelectric focusing, polarity: hydrophobic interaction chromatography, size: -gel filtration chromatography. Sds-page: binding specificity: affinity chromatography, only functional protein. Secondary: local spatial arrangement of a polypeptide backbone atoms through h-bond interactions. Quaternary: spatial arrangement of diff polypeptide chains in a protein: 1 polypeptide can be cleaved to form more than 1 protein. 1 gene can be transcribed into different mrna"s. 1 mrna can be translated into multiple proteins. All purification methods must be based on the properties of the target proteins. If the properties of the protein is unknown, different strategies have to be tried. Then electrostatic interactions bn protein molecules make proteins precipitate: initially all are soluble in buffer, add salt to solution, unwanted protein precipitates (mb remove by centrifugation) *proteins are least soluble at pi, it"s a ph thing.
