ACS BIOCHEMISTRY EXAM QUESTIONS AND CORRECT DETAILED ANSWERS | EVERYTHING YOU NEED TO PASS EXAM Step 1 of the Citric Acid Cycle - ✔✔✔ANSWER -Acetyl -CoA + Oxaloacetate --> Citrate Uses citrate synthase enzyme H2O --> CoA Rate -limiting Step of the Citric Acid Cycle - ✔✔✔ANSWER -Step 1 Acetyl -Coa + Oxaloacetate --> Citrate Step 2 of the Citric Acid Cycle - ✔✔✔ANSWER -Citrate < --> Isocitrate Uses aconitase enzyme H2O < --> H2O Step 3 of the Citric Acid Cycle - ✔✔✔ANSWER -Isocitrate --> α-
ketoglutarate Uses isocitrate dehydrogenase NAD(P)+ --> NAD(P)H + CO2 Step 4 of the Citric Acid Cycle - ✔✔✔ANSWER -α-ketoglutarate --> succinyl -CoA Uses α -ketoglutarate dehydrogenase complex CoA + NAD+ --> NADH + CO2 Step 5 of the Citric Acid Cycle - ✔✔✔ANSWER -Succinyl -CoA < --> Succinate Uses succinyl -CoA synthetase enzyme GDP + Pi < --> GTP + CoA Step 6 of the Citric Acid Cycle - ✔✔✔ANSWER -Succinate < --> Fumarate Uses succinate dehydrogenase FAD < --> FADH2 Step 7 of the Citric Acid Cycle - ✔✔✔ANSWER -Fumarate < --> L-
Malate Uses fumarase enzyme 1) OH - 2) H+ --> Step 8 of the Citric Acid Cycle - ✔✔✔ANSWER -L-Malate < --> Oxaloacetate Uses malate dehydrogenase enzyme NAD+ < --> NADH Henderson -Hasselbach Equation - ✔✔✔ANSWER -pH = pKa + log ([A-] / [HA]) FMOC Chemical Synthesis - ✔✔✔ANSWER -Used in synthesis of a growing amino acid chain to a polystyrene bead. FMOC is used as a protecting group on the N -terminus. 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. Elute with 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 charge matters. Involves electrodes and pH gradient. Protein stops at their pI when neutral. FDNB (1 -fluoro -2,3-dinitrobenzene) - ✔✔✔ANSWER -FDNB reacts with the N -terminus of the protein to produce a 2,4 -dinitrophenol
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