Menon_2011_Mol.Cell.Proteomics_10_M110 000042

Neurodegenerative diseases such as multiple sclerosis represent global health issues Accordingly there is an urgent need to understand the pathogenesis of this and other central nervous system disorders so that more effective therapeutics can be developed Cerebrospinal fluid is a potential source of important reporter molecules released from various cell types as a result of central nervous system pathology Here we report the development of an unbiased approach for the detection of reactive cerebrospinal fluid molecules and target brain proteins from patients with multiple sclerosis To help identify molecules that may serve as clinical biomarkers for multiple sclerosis we have biotinylated proteins present in the cerebrospinal fluid and tested their reactivity against brain homogenate as well as myelin and myelin-axolemmal complexes Proteins were separated by two-dimensional gel electrophoresis blotted onto membranes and probed separately with biotinylated unprocessed cerebrospinal fluid samples Protein spots that reacted to two or more multiple sclerosis-cerebrospinal fluids were further analyzed by matrix assisted laser desorption ionization-time-of-flight time-of-flight mass spectrometry In addition to previously reported proteins found in multiple sclerosis cerebrospinal fluid such as alphabeta crystallin enolase and 14-3-3-protein we have identified several additional molecules involved in mitochondrial and energy metabolism myelin gene expression and/or cytoskeletal organization These include aspartate aminotransferase cyclophilin-A quaking protein collapsin response mediator protein-2 ubiquitin carboxy-terminal hydrolase L1 and cofilin To further validate these findings the cellular expression pattern of collapsin response mediator protein-2 and ubiquitin carboxy-terminal hydrolase L1 were investigated in human chronic-active MS lesions by immunohistochemistry The observation that in multiple sclerosis lesions phosphorylated collapsin response mediator protein-2 was increased whereas Ubiquitin carboxy-terminal hydrolase L1 was down-regulated not only highlights the importance of these molecules in the pathology of this disease but also illustrates the use of our approach in attempting to decipher the complex pathological processes leading to multiple sclerosis and other neurodegenerative diseases.