Research question: What kind of molecule is the motor?
Vale wondered if the molecule powering vesicle movement was an ATPase, a protein that cleaves a phosphate off of ATP and uses that energy to change shape.
Using the reconstituted system, Vale set up a control and 4 experimental treatments. The control and all treatments included motility buffer + the reconstituted system (vesicles + purified microtubules + soluble proteins from axon cytoplasm).
+ + + indicates that many vesicles moved, + indicates that few vesicles moved slowly, and – indicates no movement.
Is the motor an ATPase?
Treatment
Amount of vesicle movement
Control
+ + +
AMP-PNP (5 mM)
-
Trypsin added to soluble protein fraction
-
Heat soluble proteins to 70\deg C
-
AMP-PNP (5'-adenylylimidodiphosphate) binds to ATPase enzymes, but cannot be hydrolyzed, so it inhibits all types of ATPases.
Trypsin is an enzyme that hydrolyzes peptide bonds.
70\deg C is hot enough to denature a protein from a squid that lives in an environment where temperatures never exceed 30\deg C.
Question: Which evidence from this experiment supports the conclusion that the motor is a protein? (Select all that apply.)
Group of answer choices
Treatment with a protein-digesting enzyme stops vesicle movement.
Heating the soluble protein fraction inhibits vesicle movement.
Vesicles move in the control.
AMP-PNP inhibits vesicle movement.