DEL Exercise 9-55 3. You wish to determine ine the distribution of phospholipids
ID: 210539 • Letter: D
Question
DEL Exercise 9-55 3. You wish to determine ine the distribution of phospholipids in the plasma membrane. You use of right-side out and inside out red that reacts with primary amino groups and two different phospholipases marized below (+ = reacted; -= did not react). blood cells. You treat the cells with SITS, a fluorescent dye . The results are sum- SphingolipaseSea snake venom Right side Inside out Right side Inside out Right side Inside out SITS out out out Phosphatidycholine Phosphatidyl-ethanolamine Sphingomyelin Phosphatidylamine From these results deduce the distribution of the four principal phospholipids in membranes. Is the cytosolic face of the plasma membrane the same as the exoplasmic face? a. b. Why did you use red blood cells for this experiment? Assume that you lysed red blood cells and let the membranes reform knowing that some will reform inside-out. How can you separate the right-side out membranes from the inside-out membranes? 4. 5. Using PubMed, find one current example of the use of lectins in research on mammalian cells Briefly describe the research and give the proper citation for the article.Explanation / Answer
Hi,
The reaction of enzymes and binding of dye indicates the presence of phospholipids. The right side means normal exposed membrane (exoplasmic). Inside out means the cytoplasmic membrane.
The phosphotidylcholine is present in the right side of the membrane as sea snake venom can react with it. So the arrangement would look like:
a. Right side membrane: phosphotidylcholine, Sphingolyelin
Inside of membrane: Phosphoridyl-ethanolamine, Phosphatidylamine.
The cytoplasmic fact and exoplasmic face are different.
b. Red blood cells do not have nucleus. So the membrane will have the lipids from cell membrane only.
4. The cells with mixture of inside out and right side membranes can be separated using chromatography. The oligosaccharides are present only in the outer membrane and also the primary amino groups are present in the inner side of the membrane. The column of affinity chromatography can be filled with beads bound to lectins. These will bind to the right side out membranes and inside out will pass throght he column. Thus they can be eluted separately.
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