Immobilization of histidine-tagged recombinant proteins onto micropatterned surfaces for cell-based functional assays

被引:51
作者
Kato, K [1 ]
Sato, H [1 ]
Iwata, H [1 ]
机构
[1] Kyoto Univ, Inst Frontier Med Sci, Sakyo Ku, Kyoto 6068507, Japan
关键词
D O I
10.1021/la050893e
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
This letter describes a method for preparing protein microarrays that allow the functional analysis of proteins at a cellular level. This method involves the utilization of recombinant proteins genetically engineered to carry a fusion tag that has an affinity for metal ions. A micropatterned alkanethiol monolayer was used to prepare a microarray having multiple spots with immobilized metal ions. The fusion protein was chelated to the spots under physiological conditions. The feasibility of the method was demonstrated by culturing neural stem cells on the microarray that displayed oligohistidine-tagged epidermal growth factor.
引用
收藏
页码:7071 / 7075
页数:5
相关论文
共 29 条
[1]   Epidermal growth factor receptor-dependent regulation of integrin-mediated signaling and cell cycle entry in epithelial cells [J].
Bill, HM ;
Knudsen, B ;
Moores, SL ;
Muthuswamy, SK ;
Rao, VR ;
Brugge, JS ;
Miranti, CK .
MOLECULAR AND CELLULAR BIOLOGY, 2004, 24 (19) :8586-8599
[2]  
CACCAMO D, 1989, LAB INVEST, V60, P390
[3]   Twenty-five years of immobilized metal ion affinity chromatography: past, present and future [J].
Chaga, GS .
JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS, 2001, 49 (1-3) :313-334
[4]   Complex phenotypic assays in high-throughput screening [J].
Clemons, PA .
CURRENT OPINION IN CHEMICAL BIOLOGY, 2004, 8 (03) :334-338
[5]   Functionalizing surfaces with nickel ions for the grafting of proteins [J].
Du Roure, O ;
Debiemme-Chouvy, C ;
Malthête, J ;
Silberzan, P .
LANGMUIR, 2003, 19 (10) :4138-4143
[6]   An extracellular matrix microarray for probing cellular differentiation [J].
Flaim, CJ ;
Chien, S ;
Bhatia, SN .
NATURE METHODS, 2005, 2 (02) :119-125
[7]   STABLE CHELATING LINKAGE FOR REVERSIBLE IMMOBILIZATION OF OLIGOHISTIDINE TAGGED PROTEINS IN THE BIACORE SURFACE-PLASMON RESONANCE DETECTOR [J].
GERSHON, PD ;
KHILKO, S .
JOURNAL OF IMMUNOLOGICAL METHODS, 1995, 183 (01) :65-76
[8]  
HERMANSON GT, 1992, IMMOBILIZED AFFINITY, pCH2
[9]   Controlling protein orientation at interfaces using histidine tags: An alternative to Ni/NTA [J].
Johnson, DL ;
Martin, LL .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2005, 127 (07) :2018-2019
[10]   Extracellular matrix effects on neurosphere cell motility [J].
Kearns, SM ;
Laywell, ED ;
Kukekov, VK ;
Steindler, DA .
EXPERIMENTAL NEUROLOGY, 2003, 182 (01) :240-244