As stated in the text, adenosine deaminase deficiency can be treated by injection of the stabilized enzyme. Why might this treatment be effective, while injection of the missing enzyme is ineffective in other conditions, such as HGPRT deficiency or a deficiency of one of the mitochondrial deoxyribo- nucleoside kinases?
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- . In Figure 6-11,a. in view of the position of HPA oxidase earlier in thepathway compared to that of HA oxidase, would youexpect people with tyrosinosis to show symptoms ofalkaptonuria?b. if a double mutant could be found, would you expecttyrosinosis to be epistatic to alkaptonuria?Design a set of experiments using chimeric proteins, composed of a mitochondrial precursor protein fused to dihydrofolate reductase (DHFR), that could be used to determine how much of the precursor protein must protrude into the mitochondrial matrix in order for the matrix targeting sequence to be cleaved by the matrix processing protease.Explain the binding of methotrexate interferes with mitochondrial import of the enzyme DHFR which was engineering to carry mitochondrial targeting sequence at its N-terminus.
- The enzyme dihydrofolate reductase (DHFR) normally resides in the cytosol, and it can be imported into mitochondria by appending a mitochondrial signal sequence. However, when this modified DHFR is incubated with methotrexate, which is a substrate analog that binds tightly to the active site, the modified DHFR is no longer imported. Propose an explanation for this finding.TPCK and TLCK are irreversible inhibitors of serine proteases. One ofthese inhibits trypsin and the other chymotrypsin. Which is which? Explainyour reasoning. Suggest the effects of each of the following mutations on the physiologicalrole of chymotrypsinogen:(a) R15S(b) C1S(c) T147SWhat would be the effect on the activity of phosphofructokinase of the mutation of Asp103 to the unusual amino acid shown below? Explain in terms of actual structures of the side chains of Asp and this unusual amino acid.
- The text states that ATP is synthesized primarily by energy metabolism,whereas other nucleoside triphosphates are formed from the action of nucleoside diphosphate kinase. What additional pathway exists for GTP synthesis?Orotic aciduria is a rare hereditary disorder due to deficient orotate phosphoribosyltransferase and orotidine-5'-decarboxylase activities that are encoded by Uridylate (UMP) synthetase gene in de novo pyrimidine biosynthesis. The characteristic of this disorder is excessive excretion of orotic acid in urine. A mutation of UMP synthetase gene has been identified as R96G (at amino acid position 96, Arginine is changed to Glycine). In orotic aciduria, predict the consequences of deficient UMP synthetase by identifying which downstream enzyme missing its action. One of important molecules to be derived from pyrimidine biosynthesis is deoxycytidine triphosphate (dCTP) for DNA replication and repair. How is dCTP synthesized, include what enzymes and important intermediates in the pathway? 3. As thymidine triphosphate (dTTP) is needed for DNA replication and repair, what enzymes and intermediates are required to get dTTPVDAC1 mitochondrial protein plays an important role in human diseases and many strategies can be taken to utilize the therapeutic potential of VDAC1. what are these diseases and strategies?
- The binding of methotrexate interferes with mitochondrial import of the enzyme DHFR which was engineering to carry mitochondrial targeting sequence at its N-terminus.Mutations in the Galactose-1-Phosphate Uridyl Transferase Gene Can you use RFLP to identify this mutation?Here we utilize a novel, well-characterized, endogenous mitochondrial mutation in the ATP6 gene of Drosophila melanogaster with a nearly complete loss of ATP synthase activity. These Drosophila mutants have a missense mutation in ATP6 (G to A transition resulting in a glycine to glutamate change at position 116 in the protein), the mitochondrial gene encoding subunit 6 of the F1Fo-ATP synthase(complex V of the respiratory chain. ATP6 allows for the hydrogen ion translocation required for the rotation of the Fo motor and the production of ATP from ADP. Drosophila ATP61 mutants model human mitochondrial encephalomyopathy and demonstrate phenotypes associated with degenerative disease, including: reduced longevity, mitochondrial pathology, progressive neural dysfunction, tissue degeneration and locomotor impairment. ATP61 Drosophila mutants exhibit a stereotyped phenotypic progression that is analogous to the symptomatic progression reported for many human mitochondrial disease…