Chapter 4. Cell Fractionation

Introduction

Analyze the Data
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Analyze the Data 4-1: Cell Fractionation

Mouse liver cells were homogenized and the homogenate subjected to equilibrium density-gradient centrifugation with sucrose gradients. Fractions obtained from these gradients were assayed for marker molecules (i.e., molecules that are limited to specific organelles). The results of these assays are shown.

Mouse liver cells were homogenized and the homogenate subjected to equilibrium density-gradient centrifugation with sucrose gradients. Fractions obtained from these gradients were assayed for marker molecules (i.e., molecules that are limited to specific organelles). The results of these assays are shown in the figure.

The marker molecules have the following functions: cytochrome oxidase is an enzyme involved in the process by which ATP is formed in the complete aerobic degradation of glucose or fatty acids; ribosomal RNA forms part of the protein-synthesizing ribosomes; catalase catalyzes decomposition of hydrogen peroxide; acid phosphatase hydrolyzes monophosphoric esters at acid pH; cytidylyl transferase is involved in phospholipid biosynthesis; and amino acid permease aids in transport of amino acids across membranes.

Question

a) Name the marker molecule and give the number of the fraction that is most enriched for each of the following cell components: lysosomes; peroxisomes; mitochondria; plasma membrane; rough endoplasmic reticulum; smooth endoplasmic reticulum.

Organelle Marker molecule Enriched fraction (no.)
Lysosomes +0nT7I+GxA4RMO9zDlUlhlZ5t5Kko4k9xz9EYP1WZcTWlLVxYCzt1aTbVqZ/aY3xeIfg1NcuKxRMsawKHSrPWWuhk8BEY+NVafyh/bebgkBxwxIGxk4WNwo0oGeeqW4G93SOaJvu9ANYZOsgDPhWAg== uB1e+MAAhCu5I2ddSS7Xt4ufjG4=
Peroxisomes DOtzo5DSzY4PXkd1N2hsqz5jW2oMISyIcyw21a4Ol04B2fkzz1+hyYHntALR6OOuNuS18oS0kpHyqV8CmPr6uFnyHjX12NcSPfhzNKiWBxRpSjz+vU6u1PvGfZUtEqAZNfOP19Z/h7Zr2jSMnpe9Sw== IdBP14h+/JLifwIGALgCs3yU144=
Mitochondria i19QQXZ7MqZi4NpTAwmdGE4B2VWCruWOLZfvglI9/6HeRJN/2vIh7FuEkW+htHlQ+k1oOARaRU/pACDpiXyvIxmjP3Evi57F9UsQmnZ5tKJVhlqPyJNL5cTbC7LU2vkQZ8ecNSRZIuwZmtnqqwqySw== fUWATwVNCnwjFZzNPWSP822QW70=
Plasma membrane Ep/G15b9mUo2NUStiHiFxhKv8TAFvnA4npo4fn25qMlVYJ16vVOfgEgGwOZRls0XttFj6zcyhKaJ5TBecB568EIaRufjurw4y+cX/b1tTNkB5lNIT3/chXzMwrkwwE3qwmR7DbrGoF7EwOhiSErDhw== Ph/cLVHjnUPhYYJ826K4NQW69/8=
Rough endoplasmic reticulum Mku+mb6dx9Z7Ic2E3IydJ0kA56jMZwuNSSC0lwfhzSE0aeHFWKqiT1UDdJy7+vFocw3VYuUdw3fTrtUasTq1V0MqfMAXQhIX23sxD5vH1y6zlElg99yj6ZyXuE98EMG7x0+GepFdl6JaL3du7SUuHA== lr6y/4enr4JgcGUx+VrG+IRqbuM=
Smooth endoplasmic reticulum KhrI6JmsP0H9lay9xdi6WAsLVLXl88tcVXzLJAMFXimEecti4noX5zAqn9XQOp9nYrk6iULNaiDT+HyQpoQqM8a0pkmXm2FkLYbi9a4NYO+3XYKk0esPUyEaHaJIVLB8U/nF+AhhPvmOyOoDPd1g4g== gXb5Dxq/WyQCBCVl95qF3VOv9oM=
Table
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Question

B5k9rTPIAPo8WNqkSzP7QYYqQ5a85hn1X8i9oZLEiAj/dcOcAPnKeZmKIDJ3vLWLFU3kRH1fpy7o1AqScB1IAUuewd6xMEmujqEOkd7/lUPIoKRE/wc5UeCS1NQwxTnieCaconr1KLCf2a9zM9ipDNwiqqEarNkHApg3kTPRSCjhXJ0BnkmOG4cWxPw=
_feedback: The rough endoplasmic reticulum is denser than the smooth endoplasmic reticulum because it is found in a gradient fraction with a higher sucrose concentration (more dense solution).

Question

1BbYEAIEw/6mh9HBYorCKTwhGk0mNCF8sgbT2LAqu/Irj7d8Kt521lnuVJJpyX+jE7bCSWQviZt2RBvGBwEvo0DWrLZfCgyMM0AQFzDsQ1EbwwneTFSPoeOcLIZ8GbTlyrdAMiA9M4UojQj1OB1oS164bWM7MMgdA/sCzumKMcUotw1EL6+yKyOP9PS3/sk2feRevsVJ2g0=

Question

tn1Uen9FWQYHfUQAkHR1u+OCoWeiIJmmOTD4SdnGy4i2H/ojkg3kV7t1AY0GBx4ELs2EMVJ/ql7pSrfpEwJB1O15vTFBIEaPcbx+qBkvtbIW59xVkqnttTyBp99xF2ZLCHhMInbmrLhe9PBWXmOjxvlTQxFN2fvVvJbESA59MTKx0dgdKUEJNV6/H/GHJLkgNQAtJuPeKGv0/DPcMxCUjQFP2LSQdlYpeL3wcfl+DMNABlOtxyafeyrjFM7PL//2q7M3h/usk32RPfAYN1AJDhqk2lEF1U/TQ3KCNw==
_feedback: Addition of a detergent to the homogenate would eliminate the basis for equilibrium density-gradient centrifugation, as each organelle membrane would be solubilized by the detergent. Subjecting a detergent-treated homogenate to equilibrium density-gradient centrifugation would most likely produce a single peak at relatively low-percent sucrose that would contain all marker molecules.

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