AQUEOUS 2-PHASE METAL AFFINITY EXTRACTION OF HEME-PROTEINS

被引：79

作者：

WUENSCHELL, GE ^{[1
]}

NARANJO, E ^{[1
]}

ARNOLD, FH ^{[1
]}

机构：

[1] CALTECH,PASADENA,CA 91125

来源：

BIOPROCESS ENGINEERING | 1990年 / 5卷 / 05期

关键词：

D O I：

10.1007/BF00376226

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

An iminodiacetic acid derivative of poly(ethylene glycol) (PEG-IDA) that chelates metal cations has been synthesized and used to extract proteins in metal affinity aqueous two-phase PEG/dextran systems. With less than 1% of the PEG substituted with chelated copper, partition coefficients are shown to increase by factors of up to 37 over extraction with unsubstituted PEG. The proteins studied are preferentially extracted into the Cu(II)PEGIDA phase in proportion to the number of accessible histidine residues on their surface. The affinity contribution to partitioning is proportional to the number of exposed histidine over a very wide range. The partition coefficients of heme-containing proteins measured in the Cu(II)PEG-IDA/dextran systems increase with the pH of the extraction mixture from pH 5.5 to pH 8.0, while partition coefficients in the unsubstituted PEG/dextran systems are very nearly independent of pH. The strong pH dependence of the metalaffinity extraction can be utilized in the recovery of the extracted protein. © 1990 Springer-Verlag.

引用

页码：199 / 202

页数：4

共 13 条

[1]

Shanbhag V.P., Axelsson C.-G., Hydrophobic interaction determined by partition in aqueous two-phase systems, Eur. J. Biochem., 60, pp. 17-22, (1975)

[2]

Johansson G., Andersson M., Parameters determining affinity partitioning of yeast enzymes using polymer-bound triazine dye ligands, J. Chromatogr., 303, pp. 39-51, (1984)

[3]

Kroner H.K., Cordes A., Schelper A., Morr M., Buckmann A. R, Kula M.R., Affinity chromatography and related techniques, pp. 491-501, (1982)

[4]

Kopperschlager G., Johansson G., Affinity partitioning with polymer-bound cibacron blue F3G-A for rapid, large-scale purification of phosphofructokinase from Baker's yeast, Anal. Biochem., 124, pp. 117-124, (1982)

[5]

Takerkart G., Segard E., Monsigny M., Partition of trypsin in two-phase systems containing a diamidino- α-ω-diphenylcarboxyl poly(ethylene glycol) as competitive inhibitor of trypsin, FEES Lett., 42, pp. 218-220, (1974)

[6]

Porath J., Carlsson J., Olsson L., Belfrage G., Metal chelate affinity chromatography: A new approach to protein fractionation, Nature, 258, pp. 598-599, (1975)

[7]

Lonnerdal, Keen C.I., Metal chelate affinity chromatography of proteins, J Appl. Biochem., 4, pp. 203-208, (1982)

[8]

Sulkowski E., Purification of proteins by IMAC, Trends Biotechnol., 3, pp. 1-7, (1985)

[9]

Hemdan E.S., Zhao Y., Sulkowski E., Porath J., Surface topography of histidine residues: A facile probe by immobilized metal ion affinity chromatography, P.N.A.S., USA, 86, pp. 1881-1815, (1989)

[10]

Buckmann A.F., Morr M., Kula M.-R., Preparation of technical grade polyethylene glycol (PEG)(M<sub>r</sub> 20 000)-N<sup>6</sup>-(2-aminoethyl)-NADH by a procedure adaptable to large-scale synthesis, Biotechnology and Applied Biochemistry, 9, pp. 258-268, (1987)

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