Academic Integrity: tutoring, explanations, and feedback — we don’t complete graded work or submit on a student’s behalf.

TM-GFP Shown are confocal microscope images of cells transfected with a transmem

ID: 48350 • Letter: T

Question

TM-GFP

Shown are confocal microscope images of cells transfected with a transmembrane protein-GFP fusion (TM GFP) and immunostained with an antibody to early endosomal antigen (EEAI). The top row is a low magnification picture and the bottom row is higher magnification. TM-GFP is thought to be a cell surface protein but evidence suggests it is prominent in an intracellular compartment. What is the general strategy for testing localization of a protein of interest in a certain intracellular compartment? What did we do in this experiment? What are early endosomes and why might we suspect TM-GFP to be in early endosomes? The low magnification images show a group of cells that stain for EEA1 but only one with TM-GFP. Why is this? In the high magnification images, are TM-GFP and EEAI signals strictly colocalized? Why or why not? What is the relationship of TM-GFP with respect to the early endosome? What are the other compartments that are downstream of early endosomes? What could be the possible fates of TM-GFP in these intracellular compartments? What strategies might we use to disrupt the trafficking of TM-GFP in the various compartments? The scale bars are shown at left. Approximate the size of these bars for the low and high magnification images.

Explanation / Answer

1. We can take the information about the protein , protein sequence and then use the sequence and bioinformatics tool , based on protein function and anottation . computational tools can provide accurate and perfect localisations.