Determining the Location of Tektins within a Vital Molecular Machine
2011-04-13
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Determining the Location of Tektins within a Vital Molecular Machine
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2011-04-13
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Abstract
Flagella and cilia play crucial roles in keeping life moving, one of the most easily
recognizable flagella being a spermazoon’s tail. These remarkable molecular
machines contain a ring of 9 outer doublet microtubules (hollow tubes, 30 nm in
diameter) each composed of protofilaments (PFs). Within each microtubule is a
hyper-stable, 3-PF “ribbon” containing the integral, filamentous protein tektin. Two
hypotheses exist regarding the location of tektin: (i) the dynein-spoke-tektin
filament and (ii) the partition model: (i) suggests that tektin forms one of the three
PFs. (ii) suggests that a tektin filament lies on top of the ribbon and is not one of
the PFs. In this project ribbons were first isolated from Strongylocentrotus
purpuratus (sea urchins). Secondly, they were partially digested by several
different enzymes and sized (showing degree of digestion) by gel electrophoresis.
Electron microscopy verified that the ribbons were still partially intact. Finally,
flanking tubulin fragments still loosely associated with the tektin filament were to
be identified by mass spectrometry. By comparing these short fragments of tubulin
to their known PF structure, we would be able to deduce whether the tektin
filament binds to the sides of the flanking tubulin PFs (as in model i) or binds
along the inside surface of the ribbon (as in model ii). The optimal digestion has
yet to be achieved; future studies aim to optimize these conditions, through use of
different digestive enzymes and digestion times, and subsequently distinguish
which model is indeed a more accurate representation of tektin location.
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Additional contributors: Christna Ouche; Dr. Richard W. Linck (faculty mentor)
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Ries, Michael. (2011). Determining the Location of Tektins within a Vital Molecular Machine. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/104426.
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