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      Detection of Differential Levels of Proteins in the Urine of Patients with Endometrial Cancer: Analysis Using Two-Dimensional Gel Electrophoresis and O-Glycan Binding Lectin

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          Abstract

          Cancers can cause some proteins to be aberrantly excreted or released in the urine, which can be used as biomarkers. To screen for potential biomarkers for endometrial cancer (ECa), the urinary proteins from patients who were newly diagnosed with early stage ECa and untreated controls were separated using two-dimensional gel electrophoresis (2-DE) and followed by image analysis. The altered levels of zinc alpha-2 glycoprotein, alpha 1-acid glycoprotein, and CD59 were detected in the patients compared to the controls. In addition, the urine of the ECa patients was also found to contain relatively lower levels of a fragment of nebulin when the 2-DE separated urinary proteins were probed using champedak galactose binding (CGB) lectin. The different levels of the nebulin fragment were further validated by subjecting the urinary protein samples to CGB lectin affinity chromatography and analysis of the bound fractions by LC-MS/MS. Our data is suggestive of the potential use of the differentially expressed urinary proteins as biomarkers for ECa although this requires further extensive validation on clinically representative populations.

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          Most cited references39

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          Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing

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            Prediction, conservation analysis, and structural characterization of mammalian mucin-type O-glycosylation sites.

            O-GalNAc-glycosylation is one of the main types of glycosylation in mammalian cells. No consensus recognition sequence for the O-glycosyltransferases is known, making prediction methods necessary to bridge the gap between the large number of known protein sequences and the small number of proteins experimentally investigated with regard to glycosylation status. From O-GLYCBASE a total of 86 mammalian proteins experimentally investigated for in vivo O-GalNAc sites were extracted. Mammalian protein homolog comparisons showed that a glycosylated serine or threonine is less likely to be precisely conserved than a nonglycosylated one. The Protein Data Bank was analyzed for structural information, and 12 glycosylated structures were obtained. All positive sites were found in coil or turn regions. A method for predicting the location for mucin-type glycosylation sites was trained using a neural network approach. The best overall network used as input amino acid composition, averaged surface accessibility predictions together with substitution matrix profile encoding of the sequence. To improve prediction on isolated (single) sites, networks were trained on isolated sites only. The final method combines predictions from the best overall network and the best isolated site network; this prediction method correctly predicted 76% of the glycosylated residues and 93% of the nonglycosylated residues. NetOGlyc 3.1 can predict sites for completely new proteins without losing its performance. The fact that the sites could be predicted from averaged properties together with the fact that glycosylation sites are not precisely conserved indicates that mucin-type glycosylation in most cases is a bulk property and not a very site-specific one. NetOGlyc 3.1 is made available at www.cbs.dtu.dk/services/netoglyc.
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              Mucin-type O-glycans in human colon and breast cancer: glycodynamics and functions.

              The glycoproteins of tumour cells are often abnormal, both in structure and in quantity. In particular, the mucin-type O-glycans have several cancer-associated structures, including the T and Tn antigens, and certain Lewis antigens. These structural changes can alter the function of the cell, and its antigenic and adhesive properties, as well as its potential to invade and metastasize. Cancer-associated mucin antigens can be exploited in diagnosis and prognosis, and in the development of cancer vaccines. The activities and Golgi localization of glycosyltransferases are the basis for the glycodynamics of cancer cells, and determine the ranges and amounts of specific O-glycans produced. This review focuses on the glycosyltransferases of colon and breast cancer cells that determine the pathways of mucin-type O-glycosylation, and the proposed functional and pathological consequences of altered O-glycans.
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                Author and article information

                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                Molecular Diversity Preservation International (MDPI)
                1422-0067
                2012
                27 July 2012
                : 13
                : 8
                : 9489-9501
                Affiliations
                [1 ]Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur 50603, Malaysia; E-Mail: alanmukangwai@ 123456yahoo.com
                [2 ]Department of Obstetrics and Gynaecology, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia; E-Mail: limbk@ 123456ummc.edu.my
                [3 ]Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia; E-Mail: onnhashim@ 123456um.edu.my
                [4 ]University of Malaya Centre for Proteomics Research, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia
                Author notes
                [* ]Author to whom correspondence should be addressed; E-Mail: adawiyah@ 123456um.edu.my ; Tel.: +603-7967-4219; Fax: +603-7967-4178.
                Article
                ijms-13-09489
                10.3390/ijms13089489
                3431808
                22949810
                92109bf6-7eec-498c-a509-b5ae70310b96
                © 2012 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.

                This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license ( http://creativecommons.org/licenses/by/3.0/).

                History
                : 26 June 2012
                : 02 July 2012
                Categories
                Article

                Molecular biology
                biomarker,champedak galactose binding lectin,2-dimensional gel electrophoresis,endometrial cancer,proteomics,urine

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