Question
Beta-glucuronidase happens to be the lysosomal enzyme that when defective causes Gaucher's disease in humans. It is a devastating disorder and depending on the nature of the mutation in the gene coding for this enzyme can result in death in early childhood. A child comes into the clinic having been diagnosed with the disease and it is your task to determine the nature of the mutation in the gene coding for the beta-glucuronidase enzyme in that particular child. The only information that you know is that the child does not have any enzyme activity. In addition, you have the cDNA and genomic clones for the enzyme in your refrigerator (from your successful experiments in questions 1 and 2 above). In order to attack this problem one should first develop a series of questions concerning the possible nature of the mutation and the experiment to answer such questions. For example- a) Is the gene deleted? What experiment can you do to answer this question. b) If you find that the gene is not deleted you might then ask- is the mRNA present- What experiment will you do to answer this question? c) If you find that the message is made what experiment will you do to determine the mutation in the gene? d) Assume that you must find out the mutation in the most expeditious manner- would you change your experiment designed in c in some manner?
Explanation / Answer
Q.No a
The gene has been deleted andsStabilizing as well as recovery of existing mutant enzyme by using chaperone molecules. Making use of drug chaperones makes sense within LSDs which are caused by ancestral mutations which do not affect enzyme functionality yet cause enzyme misfolding defects, normally missense mutation or frame-shift deletions. Chaperones functionality through stabilizing misfolded mutant proteins, such that the particular stabilized conformation could get out of the particular ER, get away from proteasomal destruction as well as be delivered to the particular lysosome in which they're able to express the residual enzyme activity. Chaperones could be substrate analogues, active-site inhibitors, cofactors or effector molecules. Any time addition occurs, deletions at your website of addition are actually found.
