The figure shows E1, E2, GST-ARD1 (1-574), GST, and GST-cytohesin-1 (C-1) added
ID: 164535 • Letter: T
Question
The figure shows E1, E2, GST-ARD1 (1-574), GST, and GST-cytohesin-1 (C-1) added to proteins that can be ubiquitinated and run on a gel. The asteroid denotes heat-inactivated protein. The samples were incubated with an anti-ubiquitin antibody. This is a picture of what lab technique (native, SDS PAGE, western)? What proteins are required for ubiquitination in this experiment? Why is Lane 3 a smear? Why were GST and C-1 added to Lanes 1 and 2? What was the purpose of the heat inactivation? What type of ubiquitarian enzyme is ARD1 (El, E2, E3)?Explanation / Answer
ANSWER:
1) GST-ARD1 catalyzed formation of ubiquitinylated proteins invitro required E1 and E2
2) E1,E2,E3, GST ARD1, GST AND GST CYTOHESIN-1
3) Smearing is when you have many bands together close enough in size that you cannot distinguish between adjacent bands With beginning molecular biologists, the most likely reason for the smearing is contamination by some stray nuclease that degraded the DNA into dozens, hundreds, or even thousands of little pieces.
4) In lane 1 c-1 activity is on effect of pH and In lane 2 GST activity was assayed on the indicated templates in the presence of either 10 mM individual dNTPs.
5) Heat inactivation is done to inactivate complement, a group of proteins present in sera that are part of the immune response
6) It is a major mechanism for regulating protein functioning in eukaryotic organism