Characterization of Proteome of Human Cerebrospinal Fluid

科研成果详情

题名	Characterization of Proteome of Human Cerebrospinal Fluid
作者	Xu, Jing ; Chen, Jinzhi ; Peskind, Elaine R.; Jin, Jinghua ; Eng, Jimmy ; Pan, Catherine ; Montine, Thomas J.; Goodlett, David R.; Zhang, Jing
著作类型	图书著作
出版日期	2006
摘要	Human cerebrospinal fluid (CSF) circulates within the ventricles of the brain and surrounds the brain in the subarachnoid space (Blennow et al., 1993a,b,c). Secretion and absorption of CSF is closely regulated with an average circulating volume of 125-150 ml in an adult. Several reasons make human CSF an ideal source for identifying biomarkers for neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and dementia with Lewy bodies (DLB). These include CSF's close proximity to the site of pathology, its high availability, and the advantage of minimal ambiguities that are commonly encountered in experimental models. However, translational research using CSF to identify biomarkers in these increasingly common diseases have not been successful to date, largely due to, in our opinion, two major grounds: (1) the underlying pathogenic events are too complex to be accurately reflected in a single molecule or small group of molecules, and (2) the lack of a complete understanding of the composition of human CSF in normal persons. Recently, we utilized discovery based proteomics (Link et al., 1999)-the identification of proteins in a given sample-to address both difficulties. The three fundamental steps in discovery-based proteomics are: (1) protein harvest or isolation; (2) protein identification by mass spectrometry (MS), typically via tandem MS (MS/MS); and (3) data mining using bioinformatics tools and databases. In the first step, proteins are often separated by three different methods: two-dimensional gel electrophoresis (2-D gel) (Hochstrasser et al., 2002), liquid chromatography (LC) (Aebersold and Goodlett, 2001; Washburn et al., 2001), and more recently, "protein chips" or activated surfaces that bind proteins based on chemical characteristics (Yip and Lomas, 2002). For the second step, MS or MS/MS identification of proteins also involves three basic steps: ionization, ion separation, and detection (Gross, 2004). Most 2-D gel and protein chip proteomic protocols are coupled with matrix-assisted laser desorption ionization (MALDI). One type of protein chip-coupled MALDI is called surface enhanced laser desorption/ionization (SELDI). In contrast, the LC-based platforms are usually interfaced to MS with electronspray ionization (ESI) (Chait and Kent, 1992). ESI-MS has emerged as one of the premier methods for examining biological molecules in solution, as it permits the direct analysis of nonvolatile compounds, such as peptides, proteins, glycoproteins, phospholipids, glycolipids, and complex carbohydrates in liquid solutions, as intact molecules without derivatization or digestion. Furthermore, ESI can be interfaced with LC readily while maintaining high sensitivity (subfemtomole). Our study utilized two popular, commercially available ESI-ion trap MS systems, the LCQ and LTQ proteomic stations made by ThermoElectron. The general method we used is referred to as shotgun proteomics, which includes methods like Multidimensional Protein Identification Technology (MudPIT) and Isotope Coded Affinity Tag (ICAT), both of which use multidimensional LC and MS/MS to separate and fragment peptides for protein identification (Link et al., 1999). The main advantage of shotgun proteomics over 2-D gel electrophoresis followed by MS is higher throughput. The advantages over the SELDI method (Forde and McCutchen-Maloney, 2002) include better coverage of proteins with high molecular weight and the ability to identify proteins directly (SELDI typically identifies unique peaks only, i.e., pattern recognition; unique protein can be identified at a later stage with extensive off-chip workup). With MudPIT and an LCQ system, we were able to identify more than 300 proteins in a previous study, focused on aging related changes in human CSF (Zhang et al., 2005a). Over the last year or so, however, it has become increasingly clear that the LCQ system is associated with several limitations with one of the major ones being slow scanning speed (Yi et al., 2002; Zhang et al., 2005b), which translates into lower reproducibility of samples when analyzed multiple times and less total proteins identified as compared to MS with faster scanning speeds, for example, an LTQ- Fourier transform (FT) ion trap. Thus, in the current work with a goal of identifying as many CSF proteins in well-characterized healthy young subjects as possible, we tested the LTQ-FT system as well as the LCQ system with a better sample preparation procedure to circumvent the limitation associated with LCQ. 2006 Elsevier Inc. All rights reserved.
ISBN	01236687
语种	英语
出版单位	International Review of Neurobiology
文献类型	专著
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/101262
专题	附属第二医院_神经内科
作者单位	1.Department of Pathology, University of Washington School of Medicine, Seattle, WA 98104, United States 2.Department of Medicinal Chemistry, University of Washington School of Medicine, Seattle, WA 98104, United States 3.Psychiatry and Behavioral Sciences and VA Mental Illness Research, Education, Clinical Center, Seattle, WA 98104, United States 4.Fred Hutchinson Cancer Research Center, Seattle, WA 98104, United States 5.Department of Neurosurgery, the 2nd Affiliated Hospital, WenZhou Medical College, Zhejiang, China
推荐引用方式 GB/T 7714	Xu, Jing,Chen, Jinzhi,Peskind, Elaine R.,et al. Characterization of Proteome of Human Cerebrospinal Fluid[M],2006.