Question from NMRWiki Q&A forum Best pulse sequence for HSQC in Guanidine-HCl

		BioNMR > NMR community > News from other NMR forums
[Question from NMRWiki Q&A forum] Best pulse sequence for HSQC in Guanidine-HCl

Webservers

NMR processing:

MDD

NMR assignment:

Backbone:

Side-chains:

NOEs:

Structure from NMR restraints:

Ab initio:

Fragment-based:

Rosetta-NMR (Robetta)

Template-based:

GeNMR

I-TASSER

Refinement:

Amber

Structure from chemical shifts:

Fragment-based:

Homology-based:

Torsion angles from chemical shifts:

Preditor

TALOS

Promega- Proline

Secondary structure from chemical shifts:

CSI (via RCI server)

TALOS

MICS caps, β-turns

d2D

PECAN

Flexibility from chemical shifts:

RCI

Interactions from chemical shifts:

HADDOCK

Chemical shifts re-referencing:

NMR model quality:

NOEs, other restraints:

Chemical shifts:

RDCs:

Pseudocontact shifts:

Protein geomtery:

SAVES2 or SAVES4

Quality Control Check

NMR spectrum prediction:

FANDAS

MestReS

V-NMR

Flexibility from structure:

Backbone S2

Methyl S2

B-factor

Molecular dynamics:

Gromacs

Amber

Antechamber

Chemical shifts prediction:

From structure:

Shiftx2

Sparta+

Camshift

CH3shift- Methyl

ArShift- Aromatic

ShiftS

Proshift

PPM

CheShift-2- Cα

From sequence:

Shifty

Camcoil

Poulsen_rc_CS

Disordered proteins:

MAXOCC

Format conversion & validation:

CCPN

From NMR-STAR 3.1

Validate NMR-STAR 3.1

NMR sample preparation:

Protein disorder:

DisMeta

Protein solubility:

Isotope labeling:

Solid-state NMR:

Thread Tools

Search this Thread

Rate Thread

Display Modes

10-23-2010, 07:42 AM

nmrlearner

Senior Member

Join Date: Jan 2005

Posts: 23,732

Points: 193,617, Level: 100

Level up: 0%, 0 Points needed

Activity: 50.7%

Last Achievements

Award-Showcase

NMR Credits: 0

NMR Points: 193,617

Downloads: 0

Uploads: 0

Best pulse sequence for HSQC in Guanidine-HCl

Best pulse sequence for HSQC in Guanidine-HCl

Hello,

I would like to acquire a series of HSQC spectra of an 15N-labeled 12 kDa protein unfolding (over the course of hours) in 3M GuaHCl. I have access to a Varian 600 spectrometer with a conventional probe. Due to the high ionic strength of the sample, tuning and shimming is a little difficult. However, I can record a sort-of-decent HSQC using a WaterGate gradient enhanced WGgNhsqc sequence. I am limited to using a protein concentration of about 250 uM. I've determined the pw90 to be 12.125 at a tpwr of 63.

Using WGgNhsqc, the protein signal is decent (relative to the guanidine), but there is high phase distortion of the guanidine signal at 6.95 ppm. The guanidine signal is "split" half up and half down.

I get slightly different results using a regular gNhsqc -- the protein seems suppressed relative to the guanidine signal, and "antiphase" to it.

I am looking for ways of increasing sensitivity -- I don't mind if the HSQC takes an hour. Does anyone have any practical advise for such an experiment? Is one pulse sequence going to be significantly superior to another in this case?

Check if somebody has answered this question on NMRWiki QA forum

Did you find this post helpful?

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
[Question from NMRWiki Q&A forum] what is best pulse sequence for c13 edit noesy on vnmrj ? what is best pulse sequence for c13 edit noesy on vnmrj ? Hi What is best pulse sequence in bio pack on vnmrj for C-13 edit NOESY ? I am working with 12 kd size protein which can exist as homo dimers ( i.e total size of homo-dimer is 24 kd ). Most frequent problems we are encountering as follows !line broadening , baseline , phase correction problems ( huge signal at 2.8 ppm )in first increments and f2f3 planes huge strips errors ! Thanking you in advance !	nmrlearner	News from other NMR forums	0	01-10-2012 03:38 PM
[Question from NMRWiki Q&A forum] Looking for hsqc-based ZZ exchange bruker pulse program Looking for hsqc-based ZZ exchange bruker pulse program hi nmrwikier ! Anyone know if the hsqc-based zz exchange pulse prog developed by Farrow et al. (1994) is available on standart bruker pulse prog library and what is its name ? Many thanks !	nmrlearner	News from other NMR forums	0	07-06-2011 06:14 AM
[Question from NMRWiki Q&A forum] CT COSY pulse sequence programming CT COSY pulse sequence programming Hi, I want to make a CT-COSY pulse sequence. But I'm bother by incremented delay. When I type in the BaseLevelRelation.h this: IN=-INand add in the microprogram the delay before and after my pi pulse,it didn't work... It seems like it calculate the d28 delay from the IN28 increment, but doesn't want to apply this delay in the pulse sequence....Any idea ? Perhaps it's about some incrementation declaration or something like that ?	nmrlearner	News from other NMR forums	0	05-04-2011 05:33 PM
[NMR analysis blog] Micropost: Pulse Sequence Blues! Micropost: Pulse Sequence Blues! Now that ENC is practically around the corner, here is a link that shows how far this conference encourages the participants’ creativity: Pulse Sequence Blues! NMR can also be fun :-) https://blogger.googleusercontent.com/tracker/6953549091784501422-8328472542836523761?l=nmr-analysis.blogspot.com	nmrlearner	News from NMR blogs	0	02-23-2011 11:21 PM
[NMR analysis blog] Micropost: Pulse Sequence Blues! Micropost: Pulse Sequence Blues! Now that ENC is practically around the corner, here is a link that shows how far this conference encourages the participants’ creativity: Pulse Sequence Blues! NMR can also be fun :-) https://blogger.googleusercontent.com/tracker/6953549091784501422-8328472542836523761?l=nmr-analysis.blogspot.com	nmrlearner	News from NMR blogs	0	02-23-2011 11:18 PM
[NMR paper] A sequential HNCA NMR pulse sequence for protein backbone assignment. A sequential HNCA NMR pulse sequence for protein backbone assignment. Related Articles A sequential HNCA NMR pulse sequence for protein backbone assignment. J Magn Reson. 2001 May;150(1):100-4 Authors: Meissner A, Sørensen OW The conventional HNCA pulse sequence suffers from the ambiguity that it cannot distinguish inter- and intraresidue correlations because the one-bond and two-bond J(NC(alpha)) coupling constants are of similar magnitude. This paper presents a novel pulse sequence, sequential HNCA, that leads to a spectrum exhibiting...	nmrlearner	Journal club	0	11-19-2010 08:32 PM
[NMR paper] A novel composite 90 degrees pulse sequence which provides distortionless NMR spectra A novel composite 90 degrees pulse sequence which provides distortionless NMR spectra and suppresses without destroying the water magnetization. Related Articles A novel composite 90 degrees pulse sequence which provides distortionless NMR spectra and suppresses without destroying the water magnetization. J Magn Reson. 1998 Jul;133(1):194-9 Authors: Sodano P, Landon C, Ptak M A novel 90 degrees composite pulse sequence which allows one to record 1D and 2D NMR spectra without disturbing the water magnetization is described. A home-written...	nmrlearner	Journal club	0	11-17-2010 11:15 PM
[NMRwiki tweet] nmrwiki: how to write #nmr pulse sequence with different carrier freqs in F1 and F3 ? nmrwiki: how to write #nmr pulse sequence with different carrier freqs in F1 and F3 ? http://qa.nmrwiki.org/question/159/ nmrwiki: how to write #nmr pulse sequence with different carrier freqs in F1 and F3 ? http://qa.nmrwiki.org/question/159/ Source: NMRWiki tweets	nmrlearner	Twitter NMR	0	08-22-2010 01:49 AM

« Previous Thread | Next Thread »

Posting Rules
You may not post new threads You may not post replies You may not post attachments You may not edit your posts BB code is On Smilies are On [IMG] code is On HTML code is On Trackbacks are Off Pingbacks are Off Refbacks are Off