BMD 410 Lecture 5 study guide 2024

What is the proteome? - functional representation of the genome -entire set of proteins expressed and modified by a cell under a particular set of biochemical conditions. -not all proteins are expressed in every cell How can we analyze the proteome? - Isolate, characterize and catalog the proteins Characterizing proteins - 1. Get access (bust open cell) 2. Purify sample 3. Assess the purity of the sample 4. Determine structure by NMR or X-ray crystallography What is the purpose of cell lysis - Disrupt the cell membrane (yields a homogenate, a mixture of the cell contents, but not bound by any membrane) Why/how is differential centrifugation used? - 1. Centrifuged at low speeds (heavy pellet forms at bottom, light supernatant at top) 2. Pellet can be centrifuged again to yield another pellet -used to separate proteins What is water of solvation? - -Adding salt to solution will bring protein out of solution -the Na and Cl ions interact with water and decrease the number of stabilizing hydrogen bones between water and protein -Salting out can be used to isolate proteins of interest from different combination of proteins. Salt can be removed by - dialysis where large proteins will retain in bag and can't move through the membrane while ions move out What is chromatography? - separation of still folded proteins -uses solid and mobile phases Chromatography can use - -charge -size -affinity for ligand -solubility -hydrophobicity -therapy stability