Lovastatin Can be Used to Produce Non-Prenylated ykt6 from PC12 Cells
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Authors
Bimber, Benjamin N.
Issue Date
2003
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
Intracellular trafficking of proteins and lipids is fundamental to normal cell
function. Essential to trafficking are SNARE proteins. SNAREs on opposing
membranes interact to form SNARE complexes which bring the opposing membranes
into close proximity and induce fusion. The SNARE ykt6 is believed to be involved in
an uncharacterized transport pathway unique to neurons. Ykt6 exists in both membrane
bound and soluble unbound forms. Unlike membrane bound ykt6, soluble ykt6 cannot
participate in SNARE complex formation and therefore does not participate in vesicle
fusion. If soluble ykt6 is able to revert to a membrane bound form this could also allow
ykt6 a degree of targeting flexibility not available to SNAREs which cannot assume a
soluble form. It would also represent a novel form of control of SNARE activity.
We hypothesize that a conformational change in ykt6 caused by a prenyl moiety
results in the inability of soluble ykt6 to form SNARE complexes. In order to test this
hypothesis, we will produce and test the SNARE binding ability of non-prenylated ykt6.
This study has established a protocol for producing and purifying non-prenylated ykt6
from PC12 cells using the drug lovastatin. We have also tested both gel filtration and
GST bead binding assays to analyze SNARE complex formation; however, a successful
assay has not been conducted at this time. Future studies will continue work towards an
assay to test the SNARE binding ability of non-prenylated ykt6. These experiments will
provide tools necessary for the advancement of our understanding of the regulation of
ykt6 and neuronal transport in general.
Description
v, 24 p.
Citation
Publisher
Kalamazoo College
License
U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder.