Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry

Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry

Bibliographic Details
Main Author: Stuart, Orville Dean
Language:English
Published: University of Cincinnati / OhioLINK 2009
Subjects:
Analytical Chemistry
ICPMS
HPLC
ESI-MS
Cerebrospinal fluid
subarachnoid hemmorhage
phosphorus
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=ucin1245823953
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spelling ndltd-OhioLink-oai-etd.ohiolink.edu-ucin12458239532021-08-03T06:13:37Z Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry Stuart, Orville Dean Analytical Chemistry ICPMS HPLC ESI-MS Cerebrospinal fluid subarachnoid hemmorhage phosphorus Cerebrospinal fluid (CSF) surrounds and serves as a protective media for the brain and central nervous system (CNS). This fluid remains isolated from other biological matrices in normal bodily conditions, therefore, an in depth analysis of CSF has the potential to reveal important details and malfunctions of many diseases that plague the nervous system. Because phosphorylation of a wide variety of proteins governs the activity of biological enzymes and systems, a method for the detection of 31P in proteins found in human cerebrospinal fluid by high-performance liquid chromatography (HPLC) coupled to inductively coupled plasma mass spectrometry (ICPMS) is described. Specifically, it is of interest to compare phosphorylated proteins/peptides from patients suffering from post subarachnoid hemorrhage (SAH) arterial vasospasms against CSF from non-diseased patients. HPLC provides a way to separate many of the components of the CSF matrix, while ICPMS allows for the simultaneous detection of various elements, specifically phosphorus, at high sensitivity. Because structural elucidation is lost in the ICPMS experiment, softer ionization techniques such as Electrospray Ionization (ESI) are thus utilized for potential protein/peptide identification. Database searching software (Spectrum Mill) is then used in this study as a means of identification and validation. SEC-ICPMS confirms the presence of phosphorylated peptides within normal CSF as well as vasospastic and non-vasospastic SAH CSF samples. Non-enzymatically digested samples with ESI-MS analysis, in conjunction with Spectrum Mill database searching, indicates a variance among phosphorylated protein species across the sample batches, however, no correlations are drawn between disease types and phosphoprotein presence or absence. 2009 English text University of Cincinnati / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=ucin1245823953 http://rave.ohiolink.edu/etdc/view?acc_num=ucin1245823953 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection NDLTD
language English
sources NDLTD
topic Analytical Chemistry
ICPMS
HPLC
ESI-MS
Cerebrospinal fluid
subarachnoid hemmorhage
phosphorus
spellingShingle Analytical Chemistry
ICPMS
HPLC
ESI-MS
Cerebrospinal fluid
subarachnoid hemmorhage
phosphorus
Stuart, Orville Dean
Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry
author Stuart, Orville Dean
author_facet Stuart, Orville Dean
author_sort Stuart, Orville Dean
title Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry
title_short Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry
title_full Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry
title_fullStr Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry
title_full_unstemmed Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry
title_sort investigation of phosphorylated proteins and peptides in human cerebrospinal fluid via high-performance liquid chromatography coupled to elemental and molecular mass spectrometry
publisher University of Cincinnati / OhioLINK
publishDate 2009
url http://rave.ohiolink.edu/etdc/view?acc_num=ucin1245823953
work_keys_str_mv AT stuartorvilledean investigationofphosphorylatedproteinsandpeptidesinhumancerebrospinalfluidviahighperformanceliquidchromatographycoupledtoelementalandmolecularmassspectrometry
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