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Unformatted text preview: ++ 6. You are studying the transport activit y of Na K ATPase in sealed “right side out” erythrocyte ghosts. It is possible to adjust the contents of the inside of the ghosts during their preparation. Beside each substance below write INSIDE or OUTSIDE to designate whether it must be present inside or outside the ghost to maintain enzyme act ivit y. a) Na+ ______________ b) ATP ______________ ++ + c) Ouabain inhibit s the activit y o f Na K ATPase by binding to the K binding site of this enzyme. Must ouabain be on the inside of the ghost or the outside of the ++ ghost to inhibit the Na K ATPase act ivit y? 7. Beside each portion of the cell listed below, write either “inside” or “outside” to designate whether it is topologically related to the inside or the outside of the cell. Interior of lysoso me ____________________ Nuclear matrix ____________________ Cytosol ____________________ Space between the two mitochondrial membranes ____________________ 8. Midtermase is a soluble enzyme that can be iso lated from the lysoso mes of fibroblasts. The isolated enzyme has 240 amino acids and is a glycoprotein with a mo lecular weight of 36,000. An analysis of this glycoprotein showed that it was a single polypept ide chain containing 4 carbohydrate units. The mRNA encoding midtermase was iso lated and used in an in vitro translat ion/translocat ion study. Five separate tubes were set up, each containing the mRNA for this protein, ATP, GTP, radio labeled amino acids, and all other ingredients necessary to achieve translat ion of the protein. Microsomes were added to each of the tubes except tube 1 which was a control. The tubes were incubated for an appropriate time and at an appropriate temperature that would enable translat ion/translocat ion to occur. The detergent, SDS, was then added to Tubes 14, fo llo wed by the addit ion of a protease to tube 2 and a deglycosylat ing enzyme to tube 3. (The deglycosylating enzyme remo ves all of the carbohydrate from the protein.) A protease was added to Tube 5; the microsomes from that tube were De Lumen of endoplasmic reticulum ____________________ mo ...
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This note was uploaded on 05/24/2010 for the course BIS 104 taught by Professor Scholey during the Spring '08 term at UC Davis.
- Spring '08