Multipurpose peptide tags for protein isolation

被引：14

作者：

Becker, Kristian ^{[1
]}

Van Alstine, James ^{[2
]}

Bulow, Leif ^{[1
]}

机构：

[1] Lund Univ, Dept Pure & Appl Biochem, Ctr Chem & Chem Engn, SE-22100 Lund, Sweden

[2] CE Healthcare Biosci AB, SE-75184 Uppsala, Sweden

来源：

JOURNAL OF CHROMATOGRAPHY A | 2008年 / 1202卷 / 01期

关键词：

green fluorescent protein; hemoglobin; lactate dehydrogenase; aqueous two-phase system; hydrophobic interaction chromatography; immobilized metal-ion affinity; chromatography;

D O I：

10.1016/j.chroma.2008.06.045

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A multifunctional peptide tag (HYDHYD) consisting of histidine, tyrosine and aspartate residues was fused to the N-terminal ends of green fluorescent protein (GFP), lactate dehydrogenase (LDH) and human hemoglobin (Hb), proteins which were subjected to ion-exchange chromatography (IEC), aqueous two-phase system partition, immobilized metal-ion affinity chromatography (IMAC), and hydrophobic interaction chromatography (HIC). Tagged GFP was retained significantly longer (>1 column volume) in both HIC and IEC. It exhibited 3 x greater partition in favor of the hydrophobic phase in a two-phase system and 96% could be bound to an IMAC column which did not bind native GFP. (C) 2008 Elsevier B.V. All rights reserved.

引用

页码：40 / 46

页数：7

共 25 条

[1] ASAView: Database and tool for solvent accessibility representation in proteins [J].

Ahmad, S ;

Gromiha, M ;

Fawareh, H ;

Sarai, A .

BMC BIOINFORMATICS, 2004, 5 (1)

[2]

ALBERTSSON PA, 1986, PARTITION CELL PARTI, P56

[3] Correlation for the partition behavior of proteins in aqueous two-phase systems: Effect of surface hydrophobicity and charge [J].

Andrews, BA ;

Schmidt, AS ;

Asenjo, JA .

BIOTECHNOLOGY AND BIOENGINEERING, 2005, 90 (03) :380-390

[4]

BECKER K, J MOL RECOG IN PRESS

[5] Partitioning of peptides and recombinant protein-peptide fusions in thermoseparating aqueous two-phase systems:: effect of peptide primary structure [J].

Berggren, K ;

Nilsson, A ;

Johansson, G ;

Bandmann, N ;

Nygren, PÅ ;

Tjerneld, F .

JOURNAL OF CHROMATOGRAPHY B, 2000, 743 (1-2) :295-306

[6] The surface exposed amino acid residues of monomeric proteins determine the partitioning in aqueous two-phase systems [J].

Berggren, K ;

Wolf, A ;

Asenjo, JA ;

Andrews, BA ;

Tjerneld, F .

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 2002, 1596 (02) :253-268

[7] ZINC IONS BOUND TO CHIMERIC HIS4 LACTATE-DEHYDROGENASE FACILITATE DECARBOXYLATION OF OXALOACETATE [J].

CARLSSON, H ;

PRACHAYASITTIKUL, V ;

BULOW, L .

PROTEIN ENGINEERING, 1993, 6 (08) :907-911

[8]

COLETTA M, 1988, J BIOL CHEM, V263, P18286

[9] Designing new metal affinity peptides by random mutagenesis of a natural metal-binding site [J].

Enzelberger, MM ;

Minning, S ;

Schmid, RD .

JOURNAL OF CHROMATOGRAPHY A, 2000, 898 (01) :83-94

[10] N-Terminal tagged lactate dehydrogenase proteins:: evaluation of relative hydrophobicity by hydrophobic interaction chromatography and aqueous two-phase system partition [J].

Fexby, S ;

Ihre, H ;

Van Alstine, J ;

Bülow, L .

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2004, 807 (01) :25-31

← 1 2 3 →