ACS Biochemistry First Semester Complete Solution Rated A 2024

Buffer Order and Henderson Hasselbach - Water, Buffer, Salt, Base pH = pKa + log(A/HA) pKa decrease as temp increase so pH decrease Ramachandran plots - Φ = N-R ψ = C-R optimal angle to prevent steric hinderance Protein structure of a helix, B sheet, reverse turn - a: LEMARKHQ B: VICYFTW RT: PSDNG how many amino acids in alpha turn helix - 3.6 Super secondary structure - helix turn helix parallel vs. antiparallel - parallel wedges face same way antiparallel face opposite Isoelectric point (pI) - pH at which a particular molecule carries no net electrical charge. Protein Separation Techniques (ALL) - Homogenization Salting Out Affinity Chromatgraphy Ion Exchange Size Exclusion/Gel Filtration H, S, A, I, SG He said all individuals SinG Protein Separation Techniques (H) - Homogenization disrupt cells, sonicator, centrifuge Protein Separation Techniques (S) - Salting out different proteins precipitate out at different salt concentrations Protein Separation Techniques (A) - Affinity chromatography add a tag; histidine binds; see how well there is an affinity for the group Protein Separation Techniques (I) - ion exchange separation based on charge; + proteins stick to - beads Protein Separation Techniques (SG) - Size Exclusion/Gel Filtration separation based on size Cuts (protein separation techniques) - cut out what dont need/isnt protein (use 40% concen. then keep liquid; if use 60% concen. then keep pellet) Dialysis - protein separation based on bad with pores 1. Moles in bag + moles in buffer 2. Divide moles by total volume - new concentration Electrophoresis - gels - move by size with electric current 1. Isoelectric focusing - purely pH 2. SDS page pH separation is by pI and charge in 2d Protein sequencing (who cuts after what) - Cyanogen bromide = methionine trypsin = K, and R chymotrypsin = F, M, L, W, Y IP - Immunoprecipitation with proteins 1. add antibodies specific to protein of interest to lysed cells 2. add antibody binding protein on beads 3. centrifuge and protein is on bead with antibodies 4. gel elect CO-IP - observe protein protein interaction Messelson Stalh - RATSHIT. Western Blotting - run gel - transfer to sheet - stained with radioactive antibody - autoradiogram - exposed antibody ELISA - indirect and sandwich - Enzyme linked immunosorbant assay 1.) indirect - antigen coated well - bind persons antibody - bind antigen with enzyme - bind substrate = rate of color 2.) Sandwich - antibody coated well - bind person antigen - bind 2nd antibody with enzyme - bind substrate = rate of color Mass spec - identify proteins by size MALDI - matrix assisted laser desorption ionization (BIG) ESI - electric spray ionization - charged molecules - time of flight detector Nucleobase - adenine (2), guanine (3), cytosine (3), thymine/Uracil (2) nucleoside - adenosine, guanosine, cytidine, thymidine, uridine nucleotide - ATP, ADP, AMP DNA facts - 3.4 A separates bases 10 nucleotides in one twist A form = dehydrated (right); B form = normal (right) z form = left handed Base hydrolysis RNA - separating nucleosides DNA Replication mechanism (RNA transcription) - putting two bases together with template present pyrophosphate leaves Transcription DNA-->RNA - initiation, elongation, termination copied 5' --> 3' template is 3' --> 5' DNA polymerase vs. RNA Polymerase - High fidelity vs. low fidelity Endonucleases - bind splice sites Ribosomal translation - initiation: factors bind 30S/40S -- ribosome