Related ArticlesIsotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pastoris.
Protein Expr Purif. 1998 Oct;14(1):97-103
Authors: Denton H, Smith M, Husi H, Uhrin D, Barlow PN, Batt CA, Sawyer L
beta-Lactoglobulin (beta-Lg) is the major whey protein in ruminant milk and has been implicated in the irreversible denaturation of milk proteins and its associated poor processing behavior during heat treatment. In order to help understand this behavior, as well as to facilitate other studies into the relationship between the molecular structure and its behavior in solution, we have prepared and purified 15N-labeled and 13C/15N-double-labelled beta-Lg in sufficient quantities to permit a full determination of the structure and dynamics using heteronuclear NMR spectroscopy. The overexpression of the labeled protein using the Pichia pastoris system proceeds with good yield but requires the removal of significant quantities of copurifying carbohydrate which otherwise interfere with the NMR experiments. At pH 2, the resulting material gives triple resonance NMR spectra of good quality that are consistent with a monomeric, globular protein rich in beta-sheet.
Overproduction of 15N-labeled r-RGD-hirudin in pichia pastoris for NMR studies.
Overproduction of 15N-labeled r-RGD-hirudin in pichia pastoris for NMR studies.
Overproduction of 15N-labeled r-RGD-hirudin in pichia pastoris for NMR studies.
Protein Pept Lett. 2010;17(10):1228-33
Authors: Wang J, Zhang Y, Li S, Liu X, Yan X, Song H, Yu M, Dai L, Mo W
The novel recombinant hirudin, r-RGD-hirudin, inhibits thrombin and platelet aggregation. Here, we reported over-expression of (15)N-labeled r-RGD-hirudin by Pichia pastoris in minimal medium. After extensive optimization, the yield of active r-RGD-hirudin reached ?600 mg/L when...
nmrlearner
Proteins
0
01-22-2011 01:52 PM
Production of isotopically labeled heterologous proteins in non-E. coli prokaryotic and eukaryotic cells
Production of isotopically labeled heterologous proteins in non-E. coli prokaryotic and eukaryotic cells
Abstract The preparation of stable isotope-labeled proteins is necessary for the application of a wide variety of NMR methods, to study the structures and dynamics of proteins and protein complexes. The E. coli expression system is generally used for the production of isotope-labeled proteins, because of the advantages of ease of handling, rapid growth, high-level protein production, and low cost for isotope-labeling. However, many eukaryotic proteins are not functionally expressed...
[NMR paper] Semisynthesis of a segmental isotopically labeled protein splicing precursor: NMR evi
Semisynthesis of a segmental isotopically labeled protein splicing precursor: NMR evidence for an unusual peptide bond at the N-extein-intein junction.
Related Articles Semisynthesis of a segmental isotopically labeled protein splicing precursor: NMR evidence for an unusual peptide bond at the N-extein-intein junction.
Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6397-402
Authors: Romanelli A, Shekhtman A, Cowburn D, Muir TW
Protein splicing is a posttranslational autocatalytic process in which an intervening sequence, termed an intein, is...
nmrlearner
Journal club
0
11-24-2010 09:51 PM
[NMR paper] Overexpression and purification of isotopically labeled Escherichia coli MutH for NMR
Overexpression and purification of isotopically labeled Escherichia coli MutH for NMR studies.
Related Articles Overexpression and purification of isotopically labeled Escherichia coli MutH for NMR studies.
Protein Expr Purif. 2003 Jun;29(2):252-8
Authors: Dutta A, Rao BJ, Chary KV
MutH is one of the enzymes involved in the methyl directed -GATC-based DNA repair system. We report a significantly optimized protocol to prepare isotopically (15N and/or 13C) labeled MutH in minimal medium with high yields for NMR studies. Under the various...
nmrlearner
Journal club
0
11-24-2010 09:01 PM
[NMR paper] alpha-->beta transition of beta-lactoglobulin as evidenced by heteronuclear NMR.
alpha-->beta transition of beta-lactoglobulin as evidenced by heteronuclear NMR.
Related Articles alpha-->beta transition of beta-lactoglobulin as evidenced by heteronuclear NMR.
J Mol Biol. 1998 Nov 6;283(4):731-9
Authors: Kuwata K, Hoshino M, Era S, Batt CA, Goto Y
Whereas bovine beta-lactoglobulin is a predominantly beta-sheet protein, it has a marked alpha-helical preference and can be considered to be a useful model of the alpha-->beta transition, a key issue for understanding the folding and biological function of a number of proteins....
nmrlearner
Journal club
0
11-17-2010 11:15 PM
[NMR paper] 1H-NMR spectroscopy of beta B2-crystallin from bovine eye lens. Conformation of the N
1H-NMR spectroscopy of beta B2-crystallin from bovine eye lens. Conformation of the N- and C-terminal extensions.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif Related Articles 1H-NMR spectroscopy of beta B2-crystallin from bovine eye lens. Conformation of the N- and C-terminal extensions.
Eur J Biochem. 1993 Apr 1;213(1):313-20
Authors: Carver JA, Cooper PG, Truscott RJ
1H-NMR spectroscopic studies of a 46-kDa homodimer, beta B2-crystallin,...
nmrlearner
Journal club
0
08-21-2010 11:53 PM
[NMR paper] High-level expression of uniformly 15N-labeled hen lysozyme in Pichia pastoris and id
High-level expression of uniformly 15N-labeled hen lysozyme in Pichia pastoris and identification of the site in hen lysozyme where phosphate ion binds using NMR measurements.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles High-level expression of uniformly 15N-labeled hen lysozyme in Pichia pastoris and identification of the site in hen lysozyme where phosphate ion binds using NMR measurements.
FEBS Lett. 1999 Apr 1;448(1):33-7
Authors: Mine S, Ueda T, Hashimoto Y, Tanaka Y,...
Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pa
Linear Mode
