Affiliations: Department of Neurology, Faculty of Medicine, Kyoto
University, 54 Shogoin-Kawaharacho, Sakyoku, Kyoto 606, Japan
Note: [] Department of Neurology, Faculty of Medicine, Kyoto University,
54 Shogoin-Kawaharacho, Sakyoku, Kyoto 606, Japan. Tel.: +81 75 751 3767; Fax:
+81 75 751 9541; E-mail: [email protected]
Abstract: In proteome analysis, it is necessary to separate proteins as a
first step prior to characterization. Thus, the overall performance of the
analysis depends strongly on the separation tool, which is usually
two-dimensional electrophoresis (2DE). We have utilized 2DE to begin
characterization of the complex pathologic processes in Alzheimer's disease
(AD). In the present study, we show how a reliable 2-DE database of brain
proteins in Alzheimer's disease was created, improving reproducibility by using
an immobilized pH gradient (IPG) for the first dimension gel electrophoresis.
The recent progress in this field, and future prospects in this area are also
discussed. Preparation of brain proteins into a suitable solubilized state
enabled us to separate over 1000 well-defined protein spots in each 2-DE. A
comparison of the density of the spots identified on the reference map between
the AD and control group, showed that 5 protein spots were significantly
increased, 28 spots were significantly decreased and 7 spots were specifically
detected in AD. Two spots among those significantly increased and one spot
among those significantly decreased were identified as GFAP related. It is
hoped that comparative studies to identify, quantitate, and characterize the
proteins differentially expressed in normal brain versus diseased brain will
give insight into the mechanisms of pathogenesis and allow the development of a
strategy to control both the etiology and course of the diseases.
Keywords: two-dimensional gel electrophoresis, Alzheimer's disease, database, Internet
