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25. Your friend works in a cell biology research lab. She is working she calls p125, because its molecular mass is 125 kiloDaltons. She knows that p125 is a transmembrane protein with three membrane-spanning domains. It has been previously reported that p125 interacts with three other proteins called p175, p80, and p50 (again, polyacrylamide gel). These four proteins in the cell. To determine how these proteins interact with the membrane, you perform a set of experiments in which you first lyse the cells and save some of your lysate, which you run in the input lane (labeled "I" in Figure Q25 below). The lysate is then subjected to a low-speed centrifugation so that you separate out the membrane fraction (which ends up in the pellet, "P") from the cytoplasm (which is in the supernatant, "S"). You then wash the pellet from the first extraction with a high-salt wash that does not disrupt the lipid bilayer, and save a little bit to run on the gel. After the high-salt wash, you centrifuge the pellet again. Your results are illustrated on the gel in Figure Q25. protein that on a on the basis of their apparent sizes on an SDS thought to exist as a protein complex 1st extraction after high-salt wash marker S P P 250 kD 150 kD 100 kD 75 kD 50 kD Figure Q25 (a) What types of bonds are most likely to be disrupted by a high salt wash? Explain your answer. I I 1 I | |

25. Your friend works in a cell biology research lab. She is working she calls p125, because its molecular mass is 125 kiloDaltons. She knows that p125 is a transmembrane protein with three membrane-spanning domains. It has been previously reported that p125 interacts with three other proteins called p175, p80, and p50 (again, polyacrylamide gel). These four proteins in the cell. To determine how these proteins interact with the membrane, you perform a set of experiments in which you first lyse the cells and save some of your lysate, which you run in the input lane (labeled "I" in Figure Q25 below). The lysate is then subjected to a low-speed centrifugation so that you separate out the membrane fraction (which ends up in the pellet, "P") from the cytoplasm (which is in the supernatant, "S"). You then wash the pellet from the first extraction with a high-salt wash that does not disrupt the lipid bilayer, and save a little bit to run on the gel. After the high-salt wash, you centrifuge the pellet again. Your results are illustrated on the gel in Figure Q25. protein that on a on the basis of their apparent sizes on an SDS thought to exist as a protein complex 1st extraction after high-salt wash marker S P P 250 kD 150 kD 100 kD 75 kD 50 kD Figure Q25 (a) What types of bonds are most likely to be disrupted by a high salt wash? Explain your answer. I I 1 I | |

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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25. Your friend works in a cell biology research lab. She is working she calls p125, because its molecular mass is 125 kiloDaltons. She knows that p125 is a transmembrane protein with three membrane-spanning domains. It has been previously reported that p125 interacts with three other proteins called p175, p80, and p50 (again, polyacrylamide gel). These four proteins in the cell. To determine how these proteins interact with the membrane, you perform a set of experiments in which you first lyse the cells and save some of your lysate, which you run in the input lane (labeled "I" in Figure Q25 below). The lysate is then subjected to a low-speed centrifugation so that you separate out the membrane fraction (which ends up in the pellet, "P") from the cytoplasm (which is in the supernatant, "S"). You then wash the pellet from the first extraction with a high-salt wash that does not disrupt the lipid bilayer, and save a little bit to run on the gel. After the high-salt wash, you centrifuge the pellet again. Your results are illustrated on the gel in Figure Q25. protein that on a on the basis of their apparent sizes on an SDS thought to exist as a protein complex 1st extraction after high-salt wash marker S P P 250 kD 150 kD 100 kD 75 kD 50 kD Figure Q25 (a) What types of bonds are most likely to be disrupted by a high salt wash? Explain your answer. I I 1 I | |
Transcribed Image Text:25. Your friend works in a cell biology research lab. She is working she calls p125, because its molecular mass is 125 kiloDaltons. She knows that p125 is a transmembrane protein with three membrane-spanning domains. It has been previously reported that p125 interacts with three other proteins called p175, p80, and p50 (again, polyacrylamide gel). These four proteins in the cell. To determine how these proteins interact with the membrane, you perform a set of experiments in which you first lyse the cells and save some of your lysate, which you run in the input lane (labeled "I" in Figure Q25 below). The lysate is then subjected to a low-speed centrifugation so that you separate out the membrane fraction (which ends up in the pellet, "P") from the cytoplasm (which is in the supernatant, "S"). You then wash the pellet from the first extraction with a high-salt wash that does not disrupt the lipid bilayer, and save a little bit to run on the gel. After the high-salt wash, you centrifuge the pellet again. Your results are illustrated on the gel in Figure Q25. protein that on a on the basis of their apparent sizes on an SDS thought to exist as a protein complex 1st extraction after high-salt wash marker S P P 250 kD 150 kD 100 kD 75 kD 50 kD Figure Q25 (a) What types of bonds are most likely to be disrupted by a high salt wash? Explain your answer. I I 1 I | |
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