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VCE Stuff => VCE Science => VCE Mathematics/Science/Technology => VCE Subjects + Help => VCE Chemistry => Topic started by: sam0001 on June 09, 2012, 01:47:40 pm

Title: Gel elctrophoresis amino acids.
Post by: sam0001 on June 09, 2012, 01:47:40 pm
Gutamic acid, serine, alanine and Arginine were anlaysed at a ph 7, in gel electophoresis. Which Amin acid would be expected to migrate the furthest towards the positive terminal?

And speaking of which, does anyone get Neap 2010, the question with gel elctophoresis?
Title: Re: Gel elctrophoresis amino acids.
Post by: diligent18 on June 09, 2012, 01:53:57 pm
Glutamic acid, because at a pH of 7, all the amino acids will be zwitterions.  Glutamic acid would be more negative than the other amino acids due to the COO- in the side chain. So it will be the most attracted to the positive terminal and will travel further? I don't know if that's correct.
Title: Re: Gel elctrophoresis amino acids.
Post by: charmanderp on June 09, 2012, 02:15:06 pm
The side chain would not be deprotonated or protonated at a pH of 7. The whole point of a zwitterion is that it has a neutral overall charge.

I think what would differentiate them would be their relative molecular masses.
Title: Re: Gel elctrophoresis amino acids.
Post by: oneoneoneone on June 09, 2012, 02:19:27 pm
Glutamic Acid's isoelectric point would be below 7 though (closer to 4ish) because of the acidic side chain. So at pH 7 it would have a -1 net charge.
Title: Re: Gel elctrophoresis amino acids.
Post by: aznxD on June 09, 2012, 02:20:46 pm
Glutamic Acid has a KPa ~4.1 so the side group would be deprotonated at a pH of 7, giving it an overall negative charge.
Title: Re: Gel elctrophoresis amino acids.
Post by: jasoN- on June 09, 2012, 02:21:08 pm
Uhh at pH 7, the term zwitterion is loosely used for all amino acids.
For amino acids, the C- and N- terminals will be ionised (hence why in all neutral-side chain amino acids, they'll be zwitterions)
In the case of amino acids with ionisable side chains (glutamic acid, aspartic acid, lysine, histidine, arginine), the net charge depends on the pH environment.
Glutamic acid will have an ionised side group (COO-) at pH 7 and will have a net charge of -1, hence it will repel the furthest away from the negative anode (i.e. closer to positive).

Edit: zwitterions only truly occur at the isoelectric point for amino acids (you don't need to know, but the isoelectric point occurs when the amino acid ceases to migrate in a pH gradient.)
For example, glutamic acid has an isoelectric point of about 3.22, that means that the overall charge will be 0 at a pH of 3.22
Title: Re: Gel elctrophoresis amino acids.
Post by: Destiny on June 09, 2012, 02:25:49 pm
Wait is the side chain charged or not charged at pH=7?  I'm getting confused, are you guys contradicting each other?
Title: Re: Gel elctrophoresis amino acids.
Post by: sam0001 on June 09, 2012, 02:32:24 pm
Lol, I have been trying to make sense of neap 2010 for about two weeks now. It just isn't clicking. I don't understand how serine could migrate further than glutamic acid??
Title: Re: Gel elctrophoresis amino acids.
Post by: charmanderp on June 09, 2012, 02:38:06 pm
Uhh at pH 7, the term zwitterion is loosely used for all amino acids.
For amino acids, the C- and N- terminals will be ionised (hence why in all neutral-side chain amino acids, they'll be zwitterions)
In the case of amino acids with ionisable side chains (glutamic acid, aspartic acid, lysine, histidine, arginine), the net charge depends on the pH environment.
Glutamic acid will have an ionised side group (COO-) at pH 7 and will have a net charge of -1, hence it will repel the furthest away from the negative anode (i.e. closer to positive).

Edit: zwitterions only truly occur at the isoelectric point for amino acids (you don't need to know, but the isoelectric point occurs when the amino acid ceases to migrate in a pH gradient.)
For example, glutamic acid has an isoelectric point of about 3.22, that means that the overall charge will be 0 at a pH of 3.22

Fair enough. This guy is most probably right, although I don't think that VCAA would then ask such a question given that we're told as part of the VCE Chemistry course that all amino acids are in their zwitterion state at pH of 7.
Title: Re: Gel elctrophoresis amino acids.
Post by: remo14 on June 09, 2012, 02:39:58 pm
don't forgot to factor in molar mass! Logically the amino acid with the lowest m/c ratio should migrate the most. Its sort of like mass spectroscopy.
Title: Re: Gel elctrophoresis amino acids.
Post by: sam0001 on June 09, 2012, 02:46:23 pm
Yeah but what I don't get with neap 2010 is the fact that glutamic acid has a greater negative charge at a ph of 6 then serine, but serine migrates further?
Title: Re: Gel elctrophoresis amino acids.
Post by: AllAboutTheLGs on June 09, 2012, 03:24:50 pm
It has something to do with the isoelectric points given in that  table
Title: Re: Gel elctrophoresis amino acids.
Post by: diligent18 on June 09, 2012, 03:44:40 pm
The side chain would not be deprotonated or protonated at a pH of 7. The whole point of a zwitterion is that it has a neutral overall charge.

I think what would differentiate them would be their relative molecular masses.
If it were based on their relative molecular masses alone then alanine should be closest to the positive electrode, which, according to answer, is incorrect. 
Also, the pH does affect the side chain.  (I've done enough practice exams with similar questions to know).
I also think that there is a high chance that a question like this may be in the exam this year.  The difficulty of the VCAA exams have been increasing year by year and they will be looking for something tricky like this to separate the excellent from the good students.
I'll probably call my Chemistry teacher now and see what he has to say about this question.
Edit: I get it now.
Title: Re: Gel elctrophoresis amino acids.
Post by: charmanderp on June 09, 2012, 03:51:25 pm
It has something to do with the isoelectric points given in that  table

What table?
Title: Re: Gel elctrophoresis amino acids.
Post by: diligent18 on June 09, 2012, 03:58:56 pm
It has something to do with the isoelectric points given in that  table

What table?
It's on the other side of the page for that question.
Title: Re: Gel elctrophoresis amino acids.
Post by: charmanderp on June 09, 2012, 04:01:21 pm
Oh right, so it actually gives a table of isoelectric points in the question? Riiiight that explains things haha, I was a bit confused
Title: Re: Gel elctrophoresis amino acids.
Post by: sam0001 on June 09, 2012, 04:44:42 pm
You can call your chem teacher? Wow.

How does it work?