Comparison of N-linked Glycoproteins in Human Whole Saliva, Parotid, Submandibular, and Sublingual Glandular Secretions Identified using Hydrazide Chemistry and Mass Spectrometry

Clinical Proteomics

Weihong Yan²  David T. Wong³  Pinmanee Boontheung²  Joseph A. Loo¹  Eric Pang²  Prasanna Ramachandran² 

[1] Department of Chemistry and Biochemistry, Molecular Biology Institute, University of California—Los Angeles, Los Angeles, USADepartment of Biological Chemistry, David Geffen School of Medicine, Los Angeles, USADepartment of Chemistry and Biochemistry, Molecular Biology Institute, University of California—Los Angeles, Los Angeles, USADepartment of Chemistry and Biochemistry, Molecular Biology Institute, University of California—Los Angeles, Los Angeles, USADepartment of Biological Chemistry, David Geffen School of Medicine, Los Angeles, USADepartment of Biological Chemistry, David Geffen School of Medicine, Los Angeles, USADepartment of Chemistry and Biochemistry, Molecular Biology Institute, University of California—Los Angeles, Los Angeles, USADepartment of Biological Chemistry, David Geffen School of Medicine, Los Angeles, USA;Department of Chemistry and Biochemistry, Molecular Biology Institute, University of California—Los Angeles, Los Angeles, USADepartment of Chemistry and Biochemistry, Molecular Biology Institute, University of California—Los Angeles, Los Angeles, USADepartment of Chemistry and Biochemistry, Molecular Biology Institute, University of California—Los Angeles, Los Angeles, USA;School of Dentistry, Dental Research Institute, University of California—Los Angeles, Los Angeles, USASchool of Dentistry, Dental Research Institute, University of California—Los Angeles, Los Angeles, USASchool of Dentistry, Dental Research Institute, University of California—Los Angeles, Los Angeles, USA

关键词: Proteomics;    Mass spectrometry;    Isoelectric focusing;    N-linked glycoproteins;    Whole saliva;    Parotid saliva;    Submandibular saliva;    Sublingual saliva;    Disease biomarker;   

DOI  :  10.1007/s12014-008-9005-0

来源: Humana Press Inc

【 摘 要 】

Abstract

Introduction

Saliva is a body fluid that holds promise for use as a diagnostic fluid for detecting diseases. Salivary proteins are known to be heavily glycosylated and are known to play functional roles in the oral cavity. We identified N-linked glycoproteins in human whole saliva, as well as the N-glycoproteins in parotid, submandibular, and sublingual glandular fluids.

Materials and Methods

We employed hydrazide chemistry to affinity enrich for N-linked glycoproteins and glycopeptides. PNGase F releases the N-peptides/proteins from the agarose-hydrazide resin, and liquid chromatography–tandem mass spectrometry was used to identify the salivary N-glycoproteins.

Results

A total of 156 formerly N-glycosylated peptides representing 77 unique N-glycoproteins were identified in salivary fluids. The total number of N-glycoproteins identified in the individual fluids was: 62, 34, 44, and 53 in whole saliva, parotid fluid, submandibular fluid, and sublingual fluid, respectively. The majority of the N-glycoproteins were annotated as extracellular proteins (40%), and several of the N-glycoproteins were annotated as membrane proteins (14%). A number of glycoproteins were differentially found in submandibular and sublingual glandular secretions.

Conclusions

Mapping the N-glycoproteome of parotid, submandibular, and sublingual saliva is important for a thorough understanding of biological processes occurring in the oral cavity and to realize the role of saliva in the overall health of human individuals. Moreover, identifying glycoproteins in saliva may also be valuable for future disease biomarker studies.

【 授权许可】

Unknown