...@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 be incredibly definitive?
Jacob
On Mon, Dec
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
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
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
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 mach...@med.unc.edu wrote:
Daniel,
You'll probably have to monitor pH changes through size changes of your
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 jin_xiec...@yahoo.com:
Dear CCP4 colleagues,
We have a protein that is composed of
]
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, Mischa
, 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, Mischa Machius
mach
]
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
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
Even without assignments, wouldn't a dramatic shift be seen
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