Screening system for D-Asp-containing proteins using D-aspartyl endopeptidase and two-dimensional gel electrophoresis.
D-Asp-containing proteins have been implicated in many aging-related diseases. To clarify the role of D-Asp-containing proteins in such diseases, we developed a screening system for these proteins using a D-aspartyl endopeptidase that specifically cleaves the proteins at the C-terminus. The digested proteins were detected by means of two-dimensional gel electrophoresis and identified using nano-liquid chromatography/tandem mass spectrometry. We were able to detect myelin basic protein, a known D-Asp-containing protein, in the brain tissues of mice; this indicates that our system is effective for screening D-Asp-containing proteins.
Sakai-Kato
Kumiko
K
Research Institute of Pharmaceutical Sciences, Musashino University, 1-1-20 Shinmachi, Nishitokyo-shi, Tokyo, 202-8585, Japan.
Kinouchi
Tadatoshi
T
Fujii
Noriko
N
Imai
Kazuhiro
K
Utsunomiya-Tate
Naoko
N
D000818:Animals; D001224:Aspartic Acid / Q000032:analysis; D016282:Aspartic Acid Endopeptidases / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D008297:Male; D051379:Mice
D001224:Aspartic Acid; D016282:Aspartic Acid Endopeptidases