Deuteration and selective labeling of alanine methyl groups of Î² 2 -adrenergic receptor expressed in a baculovirus-insect cell expression system

		BioNMR > Educational resources > Journal club
Deuteration and selective labeling of alanine methyl groups of Î² 2 -adrenergic receptor expressed in a baculovirus-insect cell expression system

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

03-08-2018, 01:24 PM

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

Deuteration and selective labeling of alanine methyl groups of Î² 2 -adrenergic receptor expressed in a baculovirus-insect cell expression system

Deuteration and selective labeling of alanine methyl groups of Î² 2 -adrenergic receptor expressed in a baculovirus-insect cell expression system

Abstract

G protein-coupled receptors (GPCRs) exist in equilibrium between multiple conformations, and their populations and exchange rates determine their functions. However, analyses of the conformational dynamics of GPCRs in lipid bilayers are still challenging, because methods for observations of NMR signals of large proteins expressed in a baculovirus-insect cell expression system (BVES) are limited. Here, we report a method to incorporate methyl-13C1H3-labeled alanine with >â??45% efficiency in highly deuterated proteins expressed in BVES. Application of the method to the NMR observations of Î²2-adrenergic receptor in micelles and in nanodiscs revealed the ligand-induced conformational differences throughout the transmembrane region of the GPCR.

Source: Journal of Biomolecular NMR

Did you find this post helpful?

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins - SelectScience Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins - SelectScience <img alt="" height="1" width="1"> Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins SelectScience Labeling schemes commonly employed for NMR investigations of high-molecular-weight proteins utilize selective incorporation of protons and 13C isotopes into methyl groups of IleÎ´1, LeuÎ´ and ValÎ³ side-chains in a highly deuterated environment (commonly ... Read here	nmrlearner	Online News	0	08-09-2017 09:26 AM
Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins - SelectScience Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins - SelectScience <img alt="" height="1" width="1"> Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins SelectScience Labeling schemes commonly employed for NMR investigations of high-molecular-weight proteins utilize selective incorporation of protons and 13C isotopes into methyl groups of IleÎ´1, LeuÎ´ and ValÎ³ side-chains in a highly deuterated environment (commonly ... Read here	nmrlearner	Online News	0	08-08-2017 10:10 AM
Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins - SelectScience Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins - SelectScience <img alt="" height="1" width="1"> Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins SelectScience Labeling schemes commonly employed for NMR investigations of high-molecular-weight proteins utilize selective incorporation of protons and 13C isotopes into methyl groups of IleÎ´1, LeuÎ´ and ValÎ³ side-chains in a highly deuterated environment (commonly ... Read here	nmrlearner	Online News	0	08-07-2017 07:31 PM
Application Note: Efficient Uniform Labeling of Proteins Expressed in Baculovirus-Infected Insect Cells Using ... - SelectScience Application Note: Efficient Uniform Labeling of Proteins Expressed in Baculovirus-Infected Insect Cells Using ... - SelectScience <img alt="" height="1" width="1"> Application Note: Efficient Uniform Labeling of Proteins Expressed in Baculovirus-Infected Insect Cells Using ... SelectScience Uniform isotope labeling is a key tool for NMR studies on recombinant proteins and their interaction with ligands of pharmaceutical interest. To be useful for most important NMR studies, a labeled protein has to be expressed with baculovirus-infected ... and more » Read here	nmrlearner	Online News	0	07-23-2017 11:38 AM
[NMR paper] Sequential protein expression and selective labeling for in-cell NMR in human cells. Sequential protein expression and selective labeling for in-cell NMR in human cells. Related Articles Sequential protein expression and selective labeling for in-cell NMR in human cells. Biochim Biophys Acta. 2015 Dec 23; Authors: Luchinat E, Secci E, Cencetti F, Bruni P Abstract BACKGROUND: In-cell NMR is a powerful technique to investigate proteins in living human cells at atomic resolution. Ideally, when studying functional processes involving protein-protein interactions by NMR, only one partner should be isotopically...	nmrlearner	Journal club	0	01-04-2016 07:49 PM
Sequential protein expression and selective labeling for in-cell NMR in human cells Sequential protein expression and selective labeling for in-cell NMR in human cells Publication date: Available online 23 December 2015 Source:Biochimica et Biophysica Acta (BBA) - General Subjects</br> Author(s): Enrico Luchinat, Erica Secci, Francesca Cencetti, Paola Bruni</br> Background In-cell NMR is a powerful technique to investigate proteins in living human cells at atomic resolution. Ideally, when studying functional processes involving protein-protein interactions by NMR, only one partner should be isotopically labeled. Here we show that...	nmrlearner	Journal club	0	12-28-2015 12:26 AM
Scrambling free combinatorial labeling of alanine-Î², isoleucine-Î´1, leucine-pro S and valine-pro S methyl groups for the detection of long range NOEs Scrambling free combinatorial labeling of alanine-Î², isoleucine-Î´1, leucine-pro S and valine-pro S methyl groups for the detection of long range NOEs Abstract Specific isotopic labeling of methyl groups in proteins has greatly extended the applicability of solution NMR spectroscopy. Simultaneous labeling of the methyl groups of several different amino acid types can offer a larger number of useful probes that can be used for structural characterisations of challenging proteins. Herein, we propose an improved AILV methyl-labeling protocol in which L...	nmrlearner	Journal club	0	11-28-2014 11:37 AM
[NMR paper] High-Resolution Heteronuclear Multidimensional NMR of Proteins in Living Insect Cells Using a Baculovirus Protein Expression System. High-Resolution Heteronuclear Multidimensional NMR of Proteins in Living Insect Cells Using a Baculovirus Protein Expression System. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--pubs.acs.org-images-pubmed-acspubs.jpg Related Articles High-Resolution Heteronuclear Multidimensional NMR of Proteins in Living Insect Cells Using a Baculovirus Protein Expression System. J Am Chem Soc. 2013 Jan 27; Authors: Hamatsu J, O'Donovan D, Tanaka T, Shirai T, Hourai Y, Mikawa T, Ikeya T, Mishima M, Boucher W, Smith BO, Laue ED, Shirakawa M, Ito Y ...	nmrlearner	Journal club	0	02-03-2013 10:19 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