#38- what was the purpose of the heat inactivation The figure shows E1, E2, GST-
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#38- what was the purpose of the heat inactivation
The figure shows E1, E2, GST-ARDI (1-574), GST, and GST-cytohesin-1 (C-1) added to proteins that can be ubiquitinated and run on a gel. The asterix denotes heat-inactivated protein. The samples were incubated with an anti-ubiquitin antibody E1 SDS-PAGE, westermy natve TE14 35, what proteins are 35. What proteins are required for ubiquitination in this experiment? 1-574 GST+ C-1 kDa OST ELE2 36, why is Lane 3 a smear? (Hint: similar to the Endo H question #16) 220- 37. Why were GST and C-1 added to Lanes 1 and 2? 90- 38. What was the purpose of the heat inactivation?Explanation / Answer
Based on the given data,
Ubiquitination is the process of breakdown of proteins with the attachment of Ubiquitin protein. This process involves three stages, namely activation, conjugation and ligation. It regulates the signaling by proteolysis.
34) The given picture shows a native Gel.
35) The presence of either E1 or E2 shows no protein in any lane; hence the E1 and E2 are required for Ubiquitination in this experiment.
36) The lane 3 is a smear, because the 1-574 inhibit the action of both E1 and E2, hence it has more protein concertation.
37) The GST and C-1 are added to the Lane 1 and 2 to check whether they can affect the action of E1 and E2. But, these proteins do not show any effect on E1 and E2.
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