Proteome profiling in tears of mycotic keratitis patients

Jevalakshmi Kandhavelu*. ChitraT, Naveen L, Lalitha P,Venkatesh Prajna N, Dharmalingam K

Author address: 

Dr.G. Venkataswamy Eye Research Institute, Aravind Medical Research Foundation, Madurai -625 020

Abstract: 

The tear film serves/performs a variety of functions and is composed of various substances, including proteins, lipids, mucins, salts, and other organic molecules. The aqueous component constitutes the majority of the tear layer, and the proteins in the tear film are believed to have a key role in the protection of the external surface from potential pathogens, and also are involved in modulation of wound healing process. Alterations in tear proteins are associated with various pathological eye conditions. Fungal keratitis caused by A. flavus is very serious and sight- threatening corneal infection which most commonly seen in patients after trauma or those with immunocompromised corneal surface. Unfortunately, early diagnosis of A. flavus keratitis still remains a big challenge. Our previous study (Ananthi et al., 2008) had already characterized the protein profile of tears from fungal keratitis by MALDI - TOF and DIGE and some proteins were found to be upregulated and some were down regulated after the infection. The aim of our study was to analyze the tear film proteins in A. flavus keratitis patients and to interpret the functions of tear proteins in A. flavus keratitis patients using Orbitrap Velos pro. Methods: Tear samples were collected from both infected patients and the healthy controls. After quantification using Bradford method multiple tear samples were pooled to reach a protein concentration of lOOjig. The tear protein content was resolved by SDS- PAGE. Subsequently, in-gel digestion was carried out and proteins were identified using LC-MS/MS. Results: SDS-PAGE analysis revealed differential expression of the proteins between control and infected tear samples. With gel-based fractionation we have identified 234 proteins in the neat tear and 834 proteins in the infected tear. Around 92 proteins from neat tear and 348 proteins from infected tear have been reported in the human tear proteome database. Remaining 142 proteins in neat tear and 469 proteins in infected tear are identified only with this study samples. Around 155 proteins are found to be common between neat and infected tear samples. In conclusion, the present study describes, the tears protein profile of A. flavus keratitis tear protein profile. Thus, we explored the influences of the disease on the tear film protein expression. This may provide insight into the pathophysiology ofA.flavus keratitis, and may lead to the identification of possible biomarkers. Acknowledgement: Department of Biotechnology, Government of India for the project and Orbitrap Mass Spectrometry facility (BT/01/CEIB/11/V1/09). 9830;Corresponding author E-mail:[email protected]
2014

abstract No: 

OP-024

Full conference title: 

Society for Indian Human and Animal Mycologists 2014
    • SIHAM