Re: [DIYbio] Protein photoswitches... photo-isomerizable chemical structures

Do you know of any work in prokaryotes?

On Fri, Jan 4, 2013 at 10:20 PM, Sebastian S. Cocioba
<scocioba@gmail.com> wrote:
> If you look into optogenetics, the channel rhodopsin pathway is very well
> understood and is being used in triggering cardio myocytes, neurons, etc
> with a pulse of light. The channel rhodopsin integration vector, tied to
> CMV, is used to transform mammalian cells but the actual ion channel came
> from "Clamy" algae cells. It has very high genetic similarity to the g
> protein linked receptors in the retina and works very well with few photons
> needed.
>
> Sebastian S Cocioba
> CEO & Founder
> New York Botanics, LLC
>
> Sent via Mobile E-Mail
>
> On Jan 4, 2013, at 7:25 PM, Nathan McCorkle <nmz787@gmail.com> wrote:
>
> So it seems adding a photoswitch isn't terribly hard, but it's an in-vitro
> operation as far as I've been reading... at least we haven't figured out the
> synBio way to do it yet.
>
> This is the seemingly simplest photo-isomerizable group:
> http://en.wikipedia.org/wiki/Azobenzene
>
> goes from trans to cis when you illuminate it, so if you add this between
> areas of a protein in the right way, you can twist the protein into or out
> of a working conformation via a pulse of light.
>
> The way to engineer a protein to receive the current in-vitro treatment is
> to identify all the surface exposed amino acids, find two that are about the
> distance relaxed or excited photoswitch, then try to engineer them to be
> cysteines and try to engineer away any other surface cysteines. Then express
> the engineered protein with your specially placed cysteine pair, purify the
> protein, add maleimide linker by using this cross-linker
> http://en.wikipedia.org/wiki/Maleimide
>
> then hope that it works, and if not try again. Here's a good slideshow of
> the concept:
>
>
> Molecular Photoswitches – Properties and Applications
> Shishi Lin, Organic Student Seminar, Yoon Group
> http://www.chem.wisc.edu/areas/organic/studsemin/lin_shishi/lin-sem.pdf
>
> It looks like we're getting close to understanding the chemistry of natural
> systems a bit better...
>
> Reversible Photocontrol of Peptide Conformation with a Rhodopsin-like
> Photoswitch
>
> http://pubs.acs.org/doi/abs/10.1021/ja301868p
>
>
>
>
>
>
> Sooo, what are some existing biological light switches that people have seen
> hacked or are ripe to be?
>
> I'm wondering if one could make a channel that spits out single nucleotides
> when illuminated, or maybe use a neuron that blebs out a vesicle with just
> one nucleotide in it. (in effort of directed DNA synthesis)
>
>
> --
> -Nathan
>
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--
-Nathan

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