scholarly journals Development of a High-Level Expression System for Deuterium-Labeled Human Serum Albumin.

2003 ◽  
Vol 53 (1) ◽  
pp. 65-69 ◽  
Author(s):  
Mihoko Tomida ◽  
Masashi Kimura ◽  
Kazuo Kuwata ◽  
Tomoya Hayashi ◽  
Yukio Okano ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Expression System ◽  
High Level Expression ◽  
High Level

scholarly journals High level expression of bikunin in Pichia pastoris by fusion of human serum albumin

AMB Express ◽  
2012 ◽  
Vol 2 (1) ◽  
pp. 14 ◽  
Author(s):  
Xing-Hua Gou ◽  
Yu-Ying Liu ◽  
Qi-Lei Chen ◽  
Jian-Jun Tang ◽  
Da-Yu Liu ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Pichia Pastoris ◽  
Serum Albumin ◽  
Human Serum ◽  
High Level Expression ◽  
High Level

The extremely high level expression of human serum albumin in the milk of transgenic mice

2012 ◽  
Vol 21 (6) ◽  
pp. 1359-1366 ◽  
Author(s):  
Xiaojie Wu ◽  
Yanli Lin ◽  
Fuyin Xiong ◽  
Yanrong Zhou ◽  
Fang Yu ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Transgenic Mice ◽  
Serum Albumin ◽  
Human Serum ◽  
High Level Expression ◽  
High Level

scholarly journals Role of Arg-410 and Tyr-411 in human serum albumin for ligand binding and esterase-like activity

2000 ◽  
Vol 349 (3) ◽  
pp. 813-819 ◽  
Author(s):  
Hiroshi WATANABE ◽  
Sumio TANASE ◽  
Keisuke NAKAJOU ◽  
Toru MARUYAMA ◽  
Ulrich KRAGH-HANSEN ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Ligand Binding ◽  
Serum Albumin ◽  
Human Serum ◽  
Aromatic Ring ◽  
Recombinant Proteins ◽  
Expression System ◽  
Guanidine Hydrochloride ◽  
Enzymic Activity ◽  
Phenolic Oxygen

Recombinant wild-type human serum albumin (rHSA), the single-residue mutants R410A, Y411A, Y411S and Y411F and the double mutant R410A/Y411A were produced using a yeast expression system. The recombinant proteins were correctly folded, as they had the same stability towards guanidine hydrochloride and the same CD spectrum as HSA isolated from serum (native HSA). Thus the global structures of the recombinant proteins are probably very similar to that of native HSA. We investigated, by ultrafiltration and CD, the high-affinity binding of two representative site II ligands, namely ketoprofen and diazepam. According to the crystal structure of HSA, the residues Arg-410 and Tyr-411 protrude into the centre of site II (in subdomain 3A), and the binding results showed that the guanidino moiety of Arg-410, the phenolic oxygen and the aromatic ring of Tyr-411 are important for ketoprofen binding. The guanidino moiety probably interacts electrostatically with the carboxy group of ketoprofen, the phenolic oxygen could make a hydrogen-bond with the keto group of the ligand, and the aromatic ring may participate in a specific stacking interaction with one of or both of the aromatic rings of ketoprofen. By contrast, Arg-410 is not important for diazepam binding. The two parts of Tyr-411 interact favourably with diazepam, and probably do so in the same way as with ketoprofen. In addition to its unique ligand binding properties, HSA also possesses an esterase-like activity, and studies with p-nitrophenyl acetate as a substrate showed that, although Arg-410 is important, the enzymic activity of HSA is much more dependent on the presence of Tyr-411. A minor activity could be registered when serine, but not alanine or phenylalanine, was present at position 411.

scholarly journals Truncated human serum albumin retains general anaesthetic binding activity

2005 ◽  
Vol 388 (1) ◽  
pp. 39-45 ◽  
Author(s):  
Renyu LIU ◽  
Jinsheng YANG ◽  
Chung-Eun HA ◽  
Nadhipuram V. BHAGAVAN ◽  
Roderic G. ECKENHOFF
Keyword(s):  
Secondary Structure ◽  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Binding Sites ◽  
Expression System ◽  
Titration Calorimetry ◽  
General Anaesthetic ◽  
Solution Studies ◽  
Domain 3

Multiple binding sites for anaesthetics in HSA (human serum albumin) make solution studies difficult to interpret. In the present study, we expressed the wild-type HSA domain 3 (wtHSAd3), a peptide with two known anaesthetic binding sites in a yeast expression system. We also expressed a site-directed mutant of domain 3 (Y411Wd3). The stability and secondary structure of the constructed fragments were determined by HX (hydrogen–tritium exchange) and CD spectroscopy. The binding of two general anaesthetics, 2-bromo-2-chloro-1,1,1-trifluoroethane and propofol, to wtHSAd3 and Y411Wd3 was determined using isothermal titration calorimetry, HX and intrinsic tryptophan fluorescence quenching. Although the expressed fragments are less stable than intact wtHSA as indicated by both CD and HX, they retain the secondary structure and anaesthetic-binding characteristics of an intact HSA molecule, but with fewer binding sites. Y411Wd3 had decreased affinity for propofol but not for 2-bromo-2-chloro-1,1,1-trifluoroethane, consistent with steric hindrance. Retention of structural features and anaesthetic binding properties with fewer binding sites in this truncated protein provide feasibility for using scaled-down models of otherwise intractable systems to gain an understanding of anaesthetic binding requirements and binding–stability relationships.

Conformational stability and warfarin-binding properties of human serum albumin studied by recombinant mutants

2001 ◽  
Vol 357 (1) ◽  
pp. 269-274 ◽  
Author(s):  
Hiroshi WATANABE ◽  
Ulrich KRAGH-HANSEN ◽  
Sumio TANASE ◽  
Keisuke NAKAJOU ◽  
Maki MITARAI ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Expression System ◽  
Conformational Stability ◽  
Affinity Binding ◽  
Binding Properties ◽  
High Affinity Binding ◽  
Yeast Expression System ◽  
High Affinity

Correctly folded recombinant wild-type human serum albumin and the single-residue mutants K199A, W214A, R218H and H242Q were produced with the use of a yeast expression system. The changes in R218H resulted in a pronounced decrease in intrinsic fluorescence. Thermodynamic parameters for thermal denaturation of the present mutants and of five additional mutants have been determined, showing small increases in stability for two mutants (R218H and H242Q) and a larger decrease in stability for one (W214A). In the last of these, denaturation was a heterogeneous process starting at physiological temperature. The high-affinity binding constant for warfarin at pH7.4 was determined by fluorescence spectroscopy: there was a significant increase in affinity for binding of warfarin to H242Q and K199A and a smaller decrease in affinity for W214A and R218H. The findings show that Trp-214 is not as essential for the high-affinity binding of warfarin as has previously been thought.

Stable Multicopy Vectors for High–Level Secretion of Recombinant Human Serum Albumin by Kluyveromyces Yeasts

1991 ◽  
Vol 9 (10) ◽  
pp. 968-975 ◽  
Author(s):  
R. Fleer ◽  
P. Yeh ◽  
N. Amellal ◽  
I. Maury ◽  
A. Fournier ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Recombinant Human Serum Albumin ◽  
High Level
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