I would like to point out that HSQC could still be applied even in such a large
protein. TROSY-HSQC has been successful in improving peaks in spectra of large
protein. Typically the sample would need to be deuterated to see the full
effect of TROSY, but even a partial deuteration can improve sig
_____
> > From: Jacob Keller [j-kell...@fsm.northwestern.edu]
> > Sent: Monday, December 06, 2010 12:54 PM
> > To: Roopa Thapar
> > Cc: CCP4BB@jiscmail.ac.uk
> > Subject: Re: [ccp4bb] pH dependent conformational change
> >
> > E
ret.
>
> It is a good experiment to try however.
>
>
> Roopa
>
>
>
>
> From: Jacob Keller [j-kell...@fsm.northwestern.edu]
> Sent: Monday, December 06, 2010 12:54 PM
> To: Roopa Thapar
> Cc: CCP4BB@jiscmail.ac.uk
> Subject: R
r
> [j-kell...@fsm.northwestern.edu]
> Sent: Monday, December 06, 2010 12:15 PM
> To: CCP4BB@JISCMAIL.AC.UK
> Subject: Re: [ccp4bb] pH dependent conformational change
>
> Wouldn't a HSQC of 15N-labeled protein be a relatively easy yes/no
> experiment? Maybe it would not
rn.edu]
> Sent: Monday, December 06, 2010 12:15 PM
> To: CCP4BB@JISCMAIL.AC.UK
> Subject: Re: [ccp4bb] pH dependent conformational change
>
> Wouldn't a HSQC of 15N-labeled protein be a relatively easy yes/no
> experiment? Maybe it would not be incredibly definitive?
>
>
]
Sent: Monday, December 06, 2010 12:15 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] pH dependent conformational change
Wouldn't a HSQC of 15N-labeled protein be a relatively easy yes/no experiment?
Maybe it would not be incredibly definitive?
Jacob
On Mon, Dec 6, 2010 at 11:10 AM, M
Hi
SAXS can be a right tool. However, how big is "short peptide linker"?
Check Nature paper by Askarieh G. and Hedhammar M. for non-His pH
sensor.
cheers
Alex
Am 06.12.2010 um 17:59 schrieb Daniel Jin :
Dear CCP4 colleagues,
We have a protein that is composed of two domains connected
Wouldn't a HSQC of 15N-labeled protein be a relatively easy yes/no
experiment? Maybe it would not be incredibly definitive?
Jacob
On Mon, Dec 6, 2010 at 11:10 AM, Mischa Machius wrote:
> Daniel,
>
> You'll probably have to monitor pH changes through size changes of your
> protein, provided the
Daniel,
You'll probably have to monitor pH changes through size changes of your
protein, provided the structural changes will indeed cause size changes.
You said "easy", so that probably rules out Small-Angle X-Ray Scattering
(SAXS), but that would be the highest-resolution method. You can try
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] pH dependent conformational change
Dear CCP4 colleagues,
We have a protein that is composed of two domains connected by a short peptide
linker. We have some indirect evidence showing that the two domains may somehow
move against each other when
Dear CCP4 colleagues,
We have a protein that is composed of two domains
connected by a short peptide linker. We have some indirect evidence showing
that the two domains may somehow move against each other when exposed to
different pH. It is unlikely to have any obvious secondary structure c
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