In-cell protein NMR and protein leakage.

		BioNMR > Educational resources > Journal club
In-cell protein NMR and protein leakage.

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:

View First Unread

Thread Tools

Search this Thread

Rate Thread

Display Modes

12-02-2010, 02:54 PM

nmrlearner

Senior Member

Join Date: Jan 2005

Posts: 23,175

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

In-cell protein NMR and protein leakage.

In-cell protein NMR and protein leakage.

In-cell protein NMR and protein leakage.

Proteins. 2010 Oct 11;

Authors: Barnes CO, Pielak GJ

In-cell nuclear magnetic resonance spectroscopy is a tool for studying proteins under physiologically relevant conditions. In some instances, however, protein signals from leaked protein are observed in the liquid surrounding the cells. Here, we examine the expression of four proteins in Escherichia coli. We describe the controls that should be used for in-cell NMR experiments and show that leakage is likely when the protein being studied exceeds ~20% of the total cellular protein. Proteins 2010. © 2010 Wiley-Liss, Inc.

PMID: 21120863 [PubMed - as supplied by publisher]

Source: PubMed

Did you find this post helpful?

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy. Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy. Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy. Biochemistry. 2011 Sep 26; Authors: Wang Q, Zhuravleva A, Gierasch LM Abstract Biology relies on functional interplay of proteins in the crowded and heterogeneous environment inside cells, and functional protein interactions are often weak and transient. Thus, methods are needed that preserve these interactions...	nmrlearner	Journal club	0	09-30-2011 06:00 AM
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy. Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy. Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy. Biochemistry. 2011 Sep 26; Authors: Wang Q, Zhuravleva A, Gierasch LM Abstract Biology relies on functional interplay of proteins in the crowded and heterogeneous environment inside cells, and functional protein interactions are often weak and transient. Thus, methods are needed that preserve these...	nmrlearner	Journal club	0	09-30-2011 05:59 AM
[NMR paper] Cell-free protein synthesis in an autoinduction system for NMR studies of protein-protein interactions. Cell-free protein synthesis in an autoinduction system for NMR studies of protein-protein interactions. Related Articles Cell-free protein synthesis in an autoinduction system for NMR studies of protein-protein interactions. J Biomol NMR. 2005 Jul;32(3):235-41 Authors: Ozawa K, Jergic S, Crowther JA, Thompson PR, Wijffels G, Otting G, Dixon NA Cell-free protein synthesis systems provide facile access to proteins in a nascent state that enables formation of soluble, native protein-protein complexes even if one of the protein components is prone...	nmrlearner	Journal club	0	12-01-2010 06:56 PM
The STINT-NMR Method for Studying In-cell Protein-Protein Interactions. The STINT-NMR Method for Studying In-cell Protein-Protein Interactions. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_120x27.gif Related Articles The STINT-NMR Method for Studying In-cell Protein-Protein Interactions. Curr Protoc Protein Sci. 2010 Aug;Chapter 17:Unit17.11 Authors: Burz DS, Shekhtman A This unit describes critical components and considerations required to study protein-protein structural interactions inside a living cell by using NMR...	nmrlearner	Journal club	0	09-05-2010 05:53 AM
[BMNRC community] In-cell NMR: three-dimensional protein structures In-cell NMR: three-dimensional protein structures http://www.nature.com/nature/journal/v458/n7234/edsumm/e090305-13.html Go to BMNRC community to find more info about this topic.	nmrlearner	News from other NMR forums	0	08-27-2010 07:58 PM
[NMR paper] Negative entropy of halothane binding to protein: 19F-NMR with a novel cell. Negative entropy of halothane binding to protein: 19F-NMR with a novel cell. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Negative entropy of halothane binding to protein: 19F-NMR with a novel cell. Biochim Biophys Acta. 1997 Mar 15;1334(2-3):117-22 Authors: Yoshida T, Tanaka M, Mori Y, Ueda I An obvious difficulty of the study of binding of volatile anesthetics to proteins is to prevent loss of the ligand during the procedure. A novel NMR tube was designed that...	nmrlearner	Journal club	0	08-22-2010 03:31 PM
[NMR paper] Negative entropy of halothane binding to protein: 19F-NMR with a novel cell. Negative entropy of halothane binding to protein: 19F-NMR with a novel cell. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Negative entropy of halothane binding to protein: 19F-NMR with a novel cell. Biochim Biophys Acta. 1997 Mar 15;1334(2-3):117-22 Authors: Yoshida T, Tanaka M, Mori Y, Ueda I An obvious difficulty of the study of binding of volatile anesthetics to proteins is to prevent loss of the ligand during the procedure. A novel NMR tube was designed that...	nmrlearner	Journal club	0	08-22-2010 03:03 PM
Cell-free protein synthesis of perdeuterated proteins for NMR studies Cell-free protein synthesis of perdeuterated proteins for NMR studies Touraj Etezady-Esfarjani, Sebastian Hiller, Cristina Villalba and Kurt Wüthrich Journal of Biomolecular NMR; 2007; 39(3); pp 229-238 Abstract: Cell-free protein synthesis protocols for uniformly deuterated proteins typically yield low, non-uniform deuteration levels. This paper introduces an E. coli cell-extract, D-S30, which enables efficient production of proteins with high deuteration levels for all non-labile hydrogen atom positions. Potential applications of the new protocol may include production of proteins...	Deano	Journal club	0	08-14-2008 10:01 PM

« 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