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 the yeast cells were culture in a fermenter. The purified (15)N-labeled r-RGD-hirudin retained full biological activity and was uniformly labeled. Heteronuclear NMR of the (15)N-labeled r-RGD-hirudin was performed for the first time, and all signals in the heteronuclear single quantum coherence (HSQC) spectrum were successfully assigned.
[NMR paper] Cell-free synthesis of 15N-labeled proteins for NMR studies.
Cell-free synthesis of 15N-labeled proteins for NMR studies.
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IUBMB Life. 2005 Sep;57(9):615-22
Authors: Ozawa K, Dixon NE, Otting G
Modern cell-free in vitro protein synthesis systems present powerful tools for the synthesis of isotope-labeled proteins in high yields. The production of selectively 15 N-labeled proteins from 15 N-labeled amino acids is particularly economic and yields are often sufficient to analyze the proteins very quickly by two-dimensional NMR...
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[NMR paper] Recombinant expression of Ole e 6, a Cys-enriched pollen allergen, in Pichia pastoris
Recombinant expression of Ole e 6, a Cys-enriched pollen allergen, in Pichia pastoris yeast: detection of partial oxidation of methionine by NMR.
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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...
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[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.
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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...
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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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11-19-2010 08:29 PM
[NMR paper] Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pa
Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pastoris.
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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...
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11-17-2010 11:15 PM
[NMR paper] Efficient enzymatic synthesis of 13C,15N-labeled DNA for NMR studies.
Efficient enzymatic synthesis of 13C,15N-labeled DNA for NMR studies.
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J Biomol NMR. 1997 Oct;10(3):245-53
Authors: Smith DE, Su JY, Jucker FM
The power of heteronuclear NMR spectroscopy to study macromolecules and their complexes has been amply demonstrated over the last decade. The obstacle to routinely applying these techniques to the study of DNA has been the synthesis of 13C,15N-labeled DNA. Here we present a simple and efficient method to generate...
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[NMR paper] Synthesis and interaction studies of 13C labeled lactone derivatives with a model pro
Synthesis and interaction studies of 13C labeled lactone derivatives with a model protein using 13C NMR.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Synthesis and interaction studies of 13C labeled lactone derivatives with a model protein using 13C NMR.
Bioorg Med Chem. 1993 Nov;1(5):389-97
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[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,...
Overproduction of 15N-labeled r-RGD-hirudin in pichia pastoris for NMR studies.
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