Channelrhodopsin-2 (ChR2) is a photoactive cation channel permeable to mono and divalent ions. The max response of wild-type ChR2 occurs at 470 nm light. ChR2 can be used to map neuronal networks. We hypothesized mutations at G181, would alter the electrostatic environment of the retinal binding pocket and alter max. Action spectra were recorded by measuring max current responses with two-electrode voltage clamp as a function of wavelength with tunable bandpass filters. G181S and G181T had little to no effect on max but had reduced function. All charged mutants had no activation, a consequence of surface expression shown by Western blotting. Therefore, we suggest that G181 is important for the proper surface expression of ChR2.

• This report represents the work of one or more WPI undergraduate students submitted to the faculty as evidence of completion of a degree requirement. WPI routinely publishes these reports on its website without editorial or peer review.

Creator

• Allen, Stephen Joseph

Publisher

• Worcester Polytechnic Institute

Identifier

• E-project-042513-150655

Advisor

• Dempski, Robert E.

Year

• 2013

Date created

• 2013-04-25

Resource type

• Major Qualifying Project

Major

• Chemistry

Rights statement

• In Copyright