Immobilization of histidine-tagged recombinant proteins onto micropatterned surfaces for cell-based functional assays.
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.
Kato
Koichi
K
Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Sato
Hideki
H
Iwata
Hiroo
H
D000818:Animals; D003585:Cell Biology / Q000295:instrumentation; D002614:Chelating Agents / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000378:metabolism; D006639:Histidine / Q000737:chemistry*; D008956:Models, Chemical; D009474:Neurons / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry; D040081:Protein Array Analysis / Q000379:methods; D051381:Rats; D011916:Rats, Inbred F344; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D013234:Stem Cells / Q000378:metabolism; D013499:Surface Properties
D002614:Chelating Agents; C471213:His-His-His-His-His-His; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D006639:Histidine; D004815:Epidermal Growth Factor