Related ArticlesRecombinant expression of Ole e 6, a Cys-enriched pollen allergen, in Pichia pastoris yeast: detection of partial oxidation of methionine by NMR.
Protein Expr Purif. 2004 Oct;37(2):336-43
Authors: Barral P, Tejera ML, Treviño MA, Batanero E, Villalba M, Bruix M, Rodríguez R
Olive pollen is one of the main causes of allergy in Mediterranean countries. Ole e 6, an olive pollen allergen, is a small (5.8 kDa) and acidic protein (pI 4.2) and no homologous proteins have been isolated or characterized so far. Ole e 6 has been efficiently expressed in the methylotrophic yeast Pichia pastoris. The cDNA encoding Ole e 6 was inserted into the plasmid vector pPIC9 and overexpressed in GS115 yeast cells. The recombinant product was purified by size-exclusion chromatography followed by reverse-phase HPLC. N-terminal sequencing, amino acid composition analysis, CD, NMR, and IgG-binding experiments were employed to characterize the purified protein. NMR data revealed the oxidation of the methionine at position 28 in approximately 50% of the recombinant protein but, although this alters its electrophoretic behavior, it did not affect folding or IgG-binding properties of rOle e 6. The recombinant form of Ole e 6 expressed in P. pastoris can be employed for structural and biochemical 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.
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...
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Proteins
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01-22-2011 01:52 PM
[NMR paper] Expression and purification of a recombinant peptide from the Alzheimer's beta-amyloi
Expression and purification of a recombinant peptide from the Alzheimer's beta-amyloid protein for solid-state NMR.
Related Articles Expression and purification of a recombinant peptide from the Alzheimer's beta-amyloid protein for solid-state NMR.
Protein Expr Purif. 2005 Jul;42(1):200-10
Authors: Sharpe S, Yau WM, Tycko R
Fibrillar protein aggregates contribute to the pathology of a number of disease states. To facilitate structural studies of these amyloid fibrils by solid-state NMR, efficient methods for the production of milligram...
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Journal club
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11-24-2010 11:14 PM
[NMR paper] NMR solution structure of Ole e 6, a major allergen from olive tree pollen.
NMR solution structure of Ole e 6, a major allergen from olive tree pollen.
Related Articles NMR solution structure of Ole e 6, a major allergen from olive tree pollen.
J Biol Chem. 2004 Sep 10;279(37):39035-41
Authors: Treviño MA, García-Mayoral MF, Barral P, Villalba M, Santoro J, Rico M, Rodríguez R, Bruix M
Ole e 6 is a pollen protein from the olive tree (Olea europaea) that exhibits allergenic activity with a high prevalence among olive-allergic individuals. The three-dimensional structure of Ole e 6 has been determined in solution by NMR...
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Journal club
0
11-24-2010 09:51 PM
[NMR paper] Expression in Pichia pastoris and characterization by circular dichroism and NMR of r
Expression in Pichia pastoris and characterization by circular dichroism and NMR of rhodostomin.
Related Articles Expression in Pichia pastoris and characterization by circular dichroism and NMR of rhodostomin.
Proteins. 2001 Jun 1;43(4):499-508
Authors: Guo RT, Chou LJ, Chen YC, Chen CY, Pari K, Jen CJ, Lo SJ, Huang SL, Lee CY, Chang TW, Chaung WJ
Rhodostomin (Rho) is a snake venom protein isolated from Calloselasma rhodostoma. Rho is a disintegrin that inhibits platelet aggregation by blocking the binding of fibrinogen to the integrin...
nmrlearner
Journal club
0
11-19-2010 08:32 PM
[NMR paper] Expression of deuterium-isotope-labelled protein in the yeast pichia pastoris for NMR
Expression of deuterium-isotope-labelled protein in the yeast pichia pastoris for NMR studies.
Related Articles Expression of deuterium-isotope-labelled protein in the yeast pichia pastoris for NMR studies.
J Biomol NMR. 2000 Aug;17(4):337-47
Authors: Morgan WD, Kragt A, Feeney J
Deuterium isotope labelling is important for NMR studies of large proteins and complexes. Many eukaryotic proteins are difficult to express in bacteria, but can be efficiently produced in the methylotrophic yeast Pichia pastoris. In order to facilitate NMR studies of...
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Journal club
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11-19-2010 08:29 PM
[NMR paper] Recombinant gene expression and 1H NMR characteristics of the kringle (2 + 3) supermo
Recombinant gene expression and 1H NMR characteristics of the kringle (2 + 3) supermodule: spectroscopic/functional individuality of plasminogen kringle domains.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--pubs.acs.org-images-acspubs.jpg Related Articles Recombinant gene expression and 1H NMR characteristics of the kringle (2 + 3) supermodule: spectroscopic/functional individuality of plasminogen kringle domains.
Biochemistry. 1996 Feb 20;35(7):2357-64
Authors: Söhndel S, Hu CK, Marti D, Affolter M, Schaller J, Llinás M, Rickli EE
...
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08-22-2010 02:27 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,...
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Journal club
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08-21-2010 04:03 PM
[Nature network NMR forum] Journal club: recombinant RNA expression (0 replies)
Journal club: recombinant RNA expression (0 replies)
Recently I was thinking about my RNA project and wishing that I could express RNA in bacteria as is typically done for recombinant proteins. I looked at the July issue of Nature Methods to see a paper describing exactly the technology I was imagining. The authors created plasmids which allow the fusion of a target RNA sequence to a tRNA scaffold which is authenically processed by the cellular machinery and protected from degradation by the intrinsic stability of the tRNA. The potential benefits of this technology for NMR of RNA...