title: On-chip identification and interaction analysis of gel-resolved proteins using a diamond-like carbon-coated plate.
abstract: We developed a novel protein chip made of a diamond-like, carbon-coated stainless steel plate (DLC plate), the surface of which is chemically modified with N-hydroxysuccinimide ester. To produce a high-density protein chip using the DLC plate, proteins separated by SDS gel electrophoresis or two-dimensional electrophoresis were electroblotted onto the DLC plate and immobilized covalently. A high blotting efficiency (25-70%) for transferring proteins from the gels onto the DLC plates was achieved by improvement of the electrophoresis device and electroblotting techniques. With the use of the DLC plate, we developed novel techniques to identify proteins immobilized on the chip and to detect protein-protein interactions on the chip by mass spectrometric analysis. We also developed a technique to identify post-translationally modified proteins, such as glycoproteins, on the protein chip.
authors.affiliation: Yokohama City University, International Graduate School of Arts and Sciences, Suchiro 1-7-29, Tsurumi, Yokohama 230-0045, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Iwafune
authors.forename: Jian-Zhong
authors.initials: JZ
authors.lastname: Tan
authors.forename: Yoko
authors.initials: Y
authors.lastname: Ino
authors.forename: Akiko
authors.initials: A
authors.lastname: Okayama
authors.forename: Yuji
authors.initials: Y
authors.lastname: Ishigaki
authors.forename: Koji
authors.initials: K
authors.lastname: Saito
authors.forename: Nobutake
authors.initials: N
authors.lastname: Suzuki
authors.forename: Mikiko
authors.initials: M
authors.lastname: Arima
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Oba
authors.forename: Shuichi
authors.initials: S
authors.lastname: Kamei
authors.forename: Michifumi
authors.initials: M
authors.lastname: Tanga
authors.forename: Takeshi
authors.initials: T
authors.lastname: Okada
authors.forename: Hisashi
authors.initials: H
authors.lastname: Hirano
mesh_terms: D000818:Animals; D002244:Carbon / Q000737:chemistry; D002417:Cattle; D018130:Diamond / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D013058:Mass Spectrometry; D040081:Protein Array Analysis / Q000379:methods*; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D013193:Stainless Steel / Q000737:chemistry*; D013388:Succinimides / Q000737:chemistry; D013499:Surface Properties
chemical_list: D006023:Glycoproteins; D011506:Proteins; D013388:Succinimides; D013193:Stainless Steel; D002244:Carbon; D018130:Diamond; C001426:N-hydroxysuccinimide