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

Ahh here is one where they're simply making a fusion protein! It's
open-access too!

LOVely enzymes – towards engineering light-controllable biocatalysts
Ulrich Krauss, Jeeyeon Lee, Stephen J. Benkovic, Karl-Erich Jaeger
http://onlinelibrary.wiley.com/doi/10.1111/j.1751-7915.2009.00140.x/abstract

On Fri, Jan 4, 2013 at 10:48 PM, Nathan McCorkle <nmz787@gmail.com> wrote:
> 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



--
-Nathan

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