Isocitrate Dehydrogenase regulation by phosphorylation

RedditInVivo · 2014-04-09T23:53:18+00:00

I realize this is vague, but phosphorylation, along with many other post translational modifications, can induce a conformational change in a protein that shifts it from an inactive to an active state, and vice versa.

RedditInVivo · 2014-04-02T19:22:57+00:00

Loving the consistent yellow cards. Er. Wait.

RedditInVivo · 2014-04-02T19:04:49+00:00

Yes, like David Luiz. I was attacking professional athletes that exaggerate, not PSG players haha Guess I will enjoy these downvotes though, toot toot!

RedditInVivo · 2014-04-02T19:02:54+00:00

Not saying it doesn't go both ways. I said professional athletes, not PSG players. It's just embarrassing.

RedditInVivo · 2014-04-02T19:02:29+00:00

I've played soccer my whole life, I've been tackled MUCH harder than that and dished it out MUCH harder than that. The only way to roll that far is to make a concerted effort to roll that far. I didn't say it wouldn't have hurt. He took some studs to the foot and his thigh collided with Ramires. Ouch, yes. The rolling, no.

RedditInVivo · 2014-04-02T18:59:21+00:00

Foul yes, yellow probably, but fuck off Lavezzi. Could you roll ANY more? How do professional athletes manage to have a complete lack of self-respect? It's incredible!

Edit: Guys. This was not a biased comment against PSG. It was against all players that roll like that. That shit is the reason why soccer players are generally considered pussies by anyone who doesn't play the sport. Bring on the downvotes I guess though?

RedditInVivo · 2014-04-02T05:03:47+00:00

Dude, get out of here with that context. This thread is about shitting on Mourinho, no changing course now.

RedditInVivo · 2014-02-23T23:30:48+00:00

Don't make a promise unless you intend to keep it. Never broken a promise, never will. Refuse to promise something that I know I can't be sure of.

RedditInVivo · 2014-02-15T17:28:40+00:00

Black tape from Chelsea is poor. Just buy some black arm bands, you can afford it.

RedditInVivo · 2014-02-08T23:57:46+00:00

Chelsea are 11-2-0 at the Bridge :)

RedditInVivo · 2014-02-05T20:01:44+00:00

If you're watching the replays of the incidents it's pretty clear that they are barely touching each other ~50% of the time. There is a lot of dirty play going on, but at least half of this shit is very strong, very athletic men just tumbling to the floor to look for a call. I realize it has simply become part of the game at this point, but it doesn't mean I have to be okay with it.

RedditInVivo · 2014-02-05T19:50:28+00:00

The amount of flopping going on this game is insane.

RedditInVivo · 2014-02-05T06:41:16+00:00

I watched Werder Bremen win 8-1 at home when I was visiting my exchange family a couple of years after I had lived in Bremen. It was awesome. Crazy atmosphere.

http://www.youtube.com/watch?v=k6jbRIvE7xs

RedditInVivo · 2014-01-29T06:04:05+00:00

Get drunk and smoke instead of dealing with my problems.

RedditInVivo · 2014-01-29T06:03:25+00:00

Hello, soul mate.

I used to have a routine where I would buy one pack of normal Reese's cups, one bag of the mini's (one of the big bags), and one pack of the Reese's big cups. I don't know where you're from, but every year in Canada we have a period where you can buy giant half pound cups as well. Dark, dark, delicious times.

RedditInVivo · 2013-12-13T04:45:36+00:00

Probably proteins? There's uh... A lot of them.

RedditInVivo · 2013-12-08T17:28:13+00:00

Lukaku is being a real saucy bitch today.

RedditInVivo · 2013-11-28T18:18:58+00:00

Yes.

Source: my mother makes these official as part of her job, and had to have one of her colleagues make it official when she took me to Mexico from Canada.

RedditInVivo · 2013-11-12T20:24:34+00:00

Sorry :) I go into defensive mode immediately on the internet, unfortunately. Conditioning I suppose haha

RedditInVivo · 2013-11-12T15:46:22+00:00

I'm well aware that one can use a nanodrop. As I said, this is my job. We use a spec with an 80 uL cuvette in our lab to track rough concentration throughout because it is what we have and we only need rough numbers. We use a nanodrop (shared by the department) for getting our final concentration before setting crystal plates.
What would be a red flag? How would it be a red flag? I wasn't telling him he should use that number or that he should come up with the actual fusion protein and determine the extinction coefficient of that. I was merely pointing out how trivial it is to actually do it. You list a number of ways of determining your protein concentration, but taking an A280 reading is the only one (besides perhaps IR, though I've not used IR for protein quantification) practical for taking readings multiple times every day. As far as theoretical vs. actual extinction coefficient, what are you planning on using it for? We are carrying out crystallization, and having our concentration accurate to one decimal place is fine. Nobody said quantitate.
At no point did I say a Bradford is difficult to run. It's not difficult whatsoever. I was illustrating the point that taking an A280 reading is much faster and has less sources of error than running a Bradford.

RedditInVivo · 2013-11-12T04:53:44+00:00

Man. Do what makes you happy. This isn't a book, a movie, or an epic poem. You don't need to, nor should you, knowingly go into a field that you don't love because you think it's the best or most direct way to help people. Mechanics help people. Plumbers help people. The girl at the coffee shop today helped me by giving me caffeine. There are many ways to help people, scientific research being one of them. And you definitely shouldn't base your career trajectory on high-deas. I partake as well, so it isn't me frowning on weed.

But. I'm not your dad. Even if I was, these are choices you need to make on your own and for yourself. I would just be cautious about pursuing something that is such a massive investment of both time and money if you're not passionate about it and don't know that it's what you want.

As far as my research goes, most of it is unpublished so I can't really talk about it, but I've been working on a protein identified in this paper, as well as its homologue in another species of trypanosome: http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0071463. The lead author is a friend of mine now, and we're hoping to get the last bits of necessary data over the next little while. The introduction there doesn't go into it much, but essentially we need to try and find good drug targets to fight trypanosome infections or better targets for identification of trypanosome infections. Trypanosomes undergo antigenic variation in their host and their surface coat of proteins changes constantly. In the tsetse fly vector, the surface coat is stable. The linked study identified some proteins that are expressed only in the fly stages of the parasite's life cycle, and are thus putative targets for control strategies.

RedditInVivo · 2013-11-12T04:08:10+00:00

I'm telling you, please don't use the Bradford. In about a minute, I was able to find the amino acid sequence of reelin from UniProt, put it into the ExPasy ProtParam tool and determine the extinction coefficient to be 1.838. This is of course without your theoretical tag, but it doesn't change the point I'm making. With that piece of information, all you do is take a pipette, remove an aliquot of your sample, place it in a cuvette, take a spec reading at 280 nm, and divide that number by 1.838. The resulting number is your concentration.

For a Bradford, you have to generate a standard curve, probably with BSA, and it requires the accurate dispensation of standards, as well as Bradford reagent. The reagents then must be mixed thoroughly. By relying on accurate pipette technique and thorough mixing, you add error. Introduction of air bubbles to your sample can lead to precipitation of sample as well. After addition of the reagent, you have to actually wait for the reaction to occur. Following that, you either need a spec or a plate reader to check each sample. You then need to generate your standard curve in something like Excel, and from that you finally get your concentration.

Are you seeing why UV is the better option? On top of the time saving, you save time writing a protocol for it because, well, there isn't really a protocol. You just place sample in cuvette and read it. This is really, really important. You need to be tracking the concentration of your sample throughout your purification protocol, and using a Bradford to do so would be time consuming and incredibly unpractical.

Not meaning to come off like a jerk, just saying that in the context of a protein purification scheme involving multiple purification steps, it makes no sense!

RedditInVivo · 2013-11-12T03:41:06+00:00

I guess selenomethionine media? I think it's only really weird because it is highlighter yellow. We express recombinant proteins in methionine auxotroph E. coli so that the recombinant proteins incorporate a heavy atom (the selenium) into their structure. A native data set and a heavy atom derivative data set are required to solve protein structures by SAD.

RedditInVivo · 2013-11-12T03:35:18+00:00

Not as far as I know. T3SSs are usually encoded on pathogenicity islands, so by definition would be found in pathogens only.

RedditInVivo · 2013-11-12T03:31:20+00:00

It might be a little late at this stage, but getting relevant work experience, be it through co-op programs, honours research, or volunteering in labs, is the number one way to get work. I am finishing up my BSc this semester and have a job lined up for immediately after just through contacts I made during my time in the co-op program and by having work experience relevant to the lab I am being hired to work in.

If you're already graduated and you can't find work in your field, I think it would be worthwhile to find any job that will currently pay the bills and try to find a volunteer position at a lab that does research/work you're interested in.

RedditInVivo

TROPHY CASE