ACS BIOCHEMISTRY EXAM QUESTIONS & ANSWERS

ACS BIOCHEMISTRY EXAM QUESTIONS & ANSWERS

Henderson-Hasselbach Equation - ANSWER pH = pKa + log ([A-] / [HA])
FMOC Chemical Synthesis - ANSWER Used in synthesis of a growing amino
acid chainto a polystyrene bead. FMOC is used as a protecting group on the Nterminus.
Salting Out (Purification) - ANSWER Changes soluble protein to solid precipitate.
Protein precipitates when the charges on the protein match the charges in the
solution.
Size-Exclusion Chromatography - ANSWER Separates sample based on size with
smaller molecules eluting later.
Ion-Exchange Chromatography - ANSWER Separates sample based on charge.
CM attracts +, DEAE attracts -. May have repulsion effect on like charges. Salt or
acid used to remove stuck proteins.
Hydrophobic/Reverse Phase Chromatography - ANSWER Beads are coated with a
carbon chain. Hydrophobic proteins stick better. Elute with non-H-bonding solvent
(acetonitrile).
Affinity Chromatography - ANSWER Attach a ligand that binds a protein to a bead.
Elutewith harsh chemicals or similar ligand.
SDS-PAGE - ANSWER Uses SDS. Gel is made from cross-linked
polyacrylamide. Separates based off of mass with smaller molecules moving
faster. Visualized with Coomassie blue.
SDS - ANSWER Sodium dodecyl sulfate. Unfolds proteins and gives them uniform
negative charge.
Isoelectric Focusing - ANSWER Variation of gel electrophoresis where protein
chargematters. Involves electrodes and pH gradient. Protein stops at their pI when
neutral.

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