In chymotrypsin, a mutant was constructed with Ser 189, which is at the bottom of the substrate-specificity pocket, changed to Lys. What effect would you predict for this Ser 189 → Lys 189 mutation?
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Chymotrypsin is a digestive enzyme that is produced in the pancreas and plays an important role in the breakdown of proteins during the digestive process. It belongs to a family of enzymes called serine proteases and is primarily responsible for cleaving peptide bonds on the carboxyl side of large hydrophobic amino acids such as phenylalanine, tryptophan, and tyrosine.
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- 1. The following are genotypes of merozygotes of E. coli with various combinations of lac operon mutations. Determine the phenotype with respect to beta-galactosidase (z), permease (y), and trans-acetylase (a) of each combination as U = uninducible, I = inducible, and C = constitutive. Choose from the following answers. i- p+ o+ z+ y- a+ / i+ p+ oc z- y- a+ Answer choices UII UIC UUU ICC CUC IUC CCU 2. The following are genotypes of merozygotes of E. coli with various combinations of lac operon mutations. Determine the phenotype with respect to beta-galactosidase (z), permease (y), and trans-acetylase (a) of each combination as U = uninducible, I = inducible, and C = constitutive. Choose from the following answers. i- p+ o+ z- y+ a+ / iS p+ o+ z- y+ a+ Answer choices ICC CCU UII IUC UIC CUC UUU2. Below, recognition sites of two of these enzymes, Pstl and Nsil, are given 5'... CTGC AG. 3... GACGTC...5 3 Pstl recognition and cleavage site 5... ATGCAT 3'... TACGTA ... 5' 3' ... Nsil recognition and cleavage site a. Does cleavage by Pstl result in a 5' or 3' overhang? What is the sequence of this overhang? b. Does cleavage by Nsil result in a 5' or 3' overhang? What is the sequence of this overhang? c. Suppose you have a cloning vector that contains a Pstl recognition site and you also have foreign DNA that was cut with Nsil. Can this DNA be ligated into the Pstl site of the vector, and if so, explain its reason and draw the new ligation site. d. Can the new ligated DNA segment sequence be cut from the vector with Pstl or Nsil ? What potential problems do you see?1. A variety of organic chelating ligands have been synthesized to tightly coordinate radioactive metal cations to identify and treat malig- nancies by coupling the metal complex to a polypeptide linker attached to a monoclonal antibody that binds specifically to a cell surface re- ceptor. One such metal complex is illustrated in the diagram on the right. The radioactive Cu-64 cation is tightly coordinated by the che- lating ligand that is, in turn, conjugated to a peptide linker attached to a monoclonal antibody. (The antibody is not shown in this diagram.) When coupled to a specifically designed monoclonal antibody, the complex binds specifically to somatostatin receptors that are ex- pressed on the surface of neuroendocrine tumors. Subsequently the entire complex with the receptor is internalized, i.e., passed into the cytoplasm, where the radioactive metal cation kills the malignant cell. CH3 HN NHNH HN NHNH HN НО НО NH HN HN HO S-S NH (a) dues in the polypeptide. Place the label…
- 3. Coronaviruses express a nucleocapsid protein that is needed for propagation, transcription, and assembly of the virus. The nucleocapsid protein must be phosphorylated by a kinase in the host cell to carry out these functions. One such kinase that has been recently reported is glycogen synthase kinase 3 or GSK-3. The following is the 10-letter sequence of the nucleocapsid protein that is recognized and phosphorylated by GSK-3: SSRGTSPARM. Note: pk. N-terminus = 9.3; pk. R = 12.5; pK. T= 13; pK. S= 13; pK. C-terminus = 4.3 a) What is the sequence of the peptide using the three-letter amino acid abbreviations? b) Draw the chemical structure of the peptide when it is at pH 8. Assign charges and label the peptide bonds. c) What is the pl of the peptide? Do not use an online resource to calculate this value. Show your work to receive credit.Scientists IPed a mutant version of KDM3A with serine 264 altered to aspartic acid (dubbed S/D mutation). They IPed S/D in cells that had not been subjected to heat shock (HS-) or heat shock (HS+). 1) what is the purpose of mutating serine-264 to aspartic acid? 2) what conclusions can scientists make based on results of lane5 and lane 6?4). Mutants unable to synthesize an essential compound G were tested with related compounds D, E, and F. The results are summarized below, where + indicates growth, and 0 indicates no growth. Mutants 1 2 3 4 D 0 0 0 Compounds E 0 INAAL F 0 0 INAY a). What is the order of gene action in the biosynthetic pathway? G
- 1. Peroxisomes are organelles surrounded by a single membrane. Soluble proteins that reside within peroxisomes are imported post-translationally, and they often contain the C-terminal tripeptide SKL (serine-lysine-leucine). a) How would you confirm that a C-terminal SKL tripeptide is a peroxisomal targeting signal? b) If cells were engineered to produce a hybrid protein consisting of a mitochondrial matrix protein (including its signal sequence) followed by a C-terminal SKL signal, where in the cell would you expect the hybrid protein to be found? Explain your reasoning.1. Complete the following table based on your lac operon simulations. Absence of lactose Presence of lactose B-galactosidase permease B-galactosidase permease I*P*O*Z*Y* Mututed ISP+O*ZY* I'P*O°Z*Y* I*P*O°ZY47. Define group transfer and its role in the cell (Hint: How are amino acids activated by aminoacyl t-RNA synthetase?).
- 5. Describe the distinct SH domains and their binding specificities. How are SH domains in regulating the specificity of siganling?1. (a) By binding one L-tryptophan molecule/monomer, the trp repressor binds to DNA to sup- press synthesis of L-tryptophan in E. coli. Below is the amino acid sequence of the helix - reverse turn - helix region of the trp repressor that binds to DNA compared to the sequence of the corresponding DNA binding motif of the Prl protein. A diagram of the trp repressor dimer is also shown. Trp Prl Trp Prl 80 -Gly-Glu-Met-Ser-Gln-Arg-Glu-Leu-Lys-Asn-Glu-Leu-Gly-Ala-Gly-Ile- -Ser-Glu-Glu-Ala-Lys-Glu-Glu-Leu-Ala-Lys-Lys-Cys-Gly-Ile-Thr-Val- trp helix 5 70 trp helix 4 Prl helix 80 Prl helix Ala-Thr-Ile-Thr-Arg-Gly-Ser-Asn-Ser-Leu-Lys-Ala-Ala- Ser-Gln-Val-Ser-Asn-Trp-Phe-Gly-Asn-Lys-Arg-Ile-Arg- reverse turn 90 Comparing the two protein sequences above, identify all amino acid pairs that differ in electrostatic charge due to proton dissociable groups (assume pH 7). Indicate the charge of both residues for each such pair. (b) Circle the pair of residues for which the electrostatic charge due to…2. The amino acid sequences in the lysozyme protein produced by the TF phage were studied by Streisinger and co-workers. One sequence is Lys-Ser-Pro-Ser-Leu-Asn-Ala. However, the result of a single nucleotide deletion and insertion of another nucleotide, this amino acid sequence changed to Lys-Val-His-His-Leu-Met-Ala. Using the codons in Table 13.1, determine the nucleotide sequences that produced a. the original amino acid sequence; and b. the subsequent changes.