Targeted Expression of TGFBIp Peptides in Mouse and Human Tissue by MALDI-Mass Spectrometry Imaging
Stromal corneal dystrophies are a group of hereditary disorders caused by mutations in the <i>TGFBI</i> gene. The mutant TGFBIp is prone to protein aggregation and the mutant protein gets deposited in the cornea, leading to severe visual impairment. The mutations lead to a corneal specif...
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doaj-0f77b15411354f43ae933b20320b7a8d2021-07-23T14:06:24ZengMDPI AGSeparations2297-87392021-07-018979710.3390/separations8070097Targeted Expression of TGFBIp Peptides in Mouse and Human Tissue by MALDI-Mass Spectrometry ImagingVenkatraman Anandalakshmi0Guillaume Hochart1David Bonnel2Jonathan Stauber3Shigeto Shimmura4Rajamani Lakshminarayanan5Konstantin Pervushin6Jodhbir S. Mehta7Singapore Eye Research Institute, The Academia, 20 College Road, Level 6 Discovery Tower, Singapore 169856, SingaporeImaBiotech, Parc Eurasanté, 152 Rue Du Docteur Yersin, 59120 Loos, FranceImaBiotech, Parc Eurasanté, 152 Rue Du Docteur Yersin, 59120 Loos, FranceImaBiotech, Parc Eurasanté, 152 Rue Du Docteur Yersin, 59120 Loos, FranceDepartment of Ophthalmology, Keio University, 35 Shinanomachi, Tokyo 160-8582, JapanSingapore Eye Research Institute, The Academia, 20 College Road, Level 6 Discovery Tower, Singapore 169856, SingaporeSchool of Biological Sciences, Nanyang Technological University, Singapore 637551, SingaporeSingapore Eye Research Institute, The Academia, 20 College Road, Level 6 Discovery Tower, Singapore 169856, SingaporeStromal corneal dystrophies are a group of hereditary disorders caused by mutations in the <i>TGFBI</i> gene. The mutant TGFBIp is prone to protein aggregation and the mutant protein gets deposited in the cornea, leading to severe visual impairment. The mutations lead to a corneal specific protein aggregation suggesting the involvement of tissue-specific factors. The exact molecular mechanism of the process of tissue-specific protein aggregation remains to be elucidated. Differential proteolysis of mutant TGFBIp is a critical component of the disease pathology. The differential proteolysis gives rise to shorter peptides that are highly aggregation-prone and initiate the aggregation cascade. Analyzing the proteolytic processing of the different TGFBIp mutant may provide insight to aid in understanding the amyloid aggregation mechanism. We developed a MALDI-MSI methodology to identify expression and spatial localization of TGFBIp peptides in the cornea. Corneal tissue samples were collected from both control and dystrophic patients (with 2 different mutations), embedded in OCT and sectioned. The sections were trypsin digested and subjected to mass spectrometry imaging using a targeted approach to detect TGFBIp. MALDI-MSI identified peptides from TGFBIp that co-localized with the amyloid corneal deposits. In addition to the relative abundance data, the specific location of the peptides across the corneal sections as molecular signatures was also identified. Spatial distribution and intensity of the TGFBIp peptides showed differences between diseased and control models but also between the two LCD phenotypes. The TGFBIp peptide with <i>m</i>/<i>z</i> of 787.474 and <i>m</i>/<i>z</i> of 1179.579 showed increased expression in both LCD mutants compared to the controls. The peptide with <i>m</i>/<i>z</i> of 929.5 showed increased expression in the LCD phenotype with H626R mutation while the peptide with <i>m</i>/<i>z</i> of 1315.802 was abundant in the sample with R124C mutation. This initial report of 2D spatial protein signature and localization of TGFBIp may be expanded to other mutations to understand the proteolytic patterns of TGFBIp in different mutations.https://www.mdpi.com/2297-8739/8/7/97amyloid fibrilscornealattice corneal dystrophymatrix-assisted laser desorption ionization (MALDI)mass spectrometry imaging (MSI)TGFBIp |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Venkatraman Anandalakshmi Guillaume Hochart David Bonnel Jonathan Stauber Shigeto Shimmura Rajamani Lakshminarayanan Konstantin Pervushin Jodhbir S. Mehta |
spellingShingle |
Venkatraman Anandalakshmi Guillaume Hochart David Bonnel Jonathan Stauber Shigeto Shimmura Rajamani Lakshminarayanan Konstantin Pervushin Jodhbir S. Mehta Targeted Expression of TGFBIp Peptides in Mouse and Human Tissue by MALDI-Mass Spectrometry Imaging Separations amyloid fibrils cornea lattice corneal dystrophy matrix-assisted laser desorption ionization (MALDI) mass spectrometry imaging (MSI) TGFBIp |
author_facet |
Venkatraman Anandalakshmi Guillaume Hochart David Bonnel Jonathan Stauber Shigeto Shimmura Rajamani Lakshminarayanan Konstantin Pervushin Jodhbir S. Mehta |
author_sort |
Venkatraman Anandalakshmi |
title |
Targeted Expression of TGFBIp Peptides in Mouse and Human Tissue by MALDI-Mass Spectrometry Imaging |
title_short |
Targeted Expression of TGFBIp Peptides in Mouse and Human Tissue by MALDI-Mass Spectrometry Imaging |
title_full |
Targeted Expression of TGFBIp Peptides in Mouse and Human Tissue by MALDI-Mass Spectrometry Imaging |
title_fullStr |
Targeted Expression of TGFBIp Peptides in Mouse and Human Tissue by MALDI-Mass Spectrometry Imaging |
title_full_unstemmed |
Targeted Expression of TGFBIp Peptides in Mouse and Human Tissue by MALDI-Mass Spectrometry Imaging |
title_sort |
targeted expression of tgfbip peptides in mouse and human tissue by maldi-mass spectrometry imaging |
publisher |
MDPI AG |
series |
Separations |
issn |
2297-8739 |
publishDate |
2021-07-01 |
description |
Stromal corneal dystrophies are a group of hereditary disorders caused by mutations in the <i>TGFBI</i> gene. The mutant TGFBIp is prone to protein aggregation and the mutant protein gets deposited in the cornea, leading to severe visual impairment. The mutations lead to a corneal specific protein aggregation suggesting the involvement of tissue-specific factors. The exact molecular mechanism of the process of tissue-specific protein aggregation remains to be elucidated. Differential proteolysis of mutant TGFBIp is a critical component of the disease pathology. The differential proteolysis gives rise to shorter peptides that are highly aggregation-prone and initiate the aggregation cascade. Analyzing the proteolytic processing of the different TGFBIp mutant may provide insight to aid in understanding the amyloid aggregation mechanism. We developed a MALDI-MSI methodology to identify expression and spatial localization of TGFBIp peptides in the cornea. Corneal tissue samples were collected from both control and dystrophic patients (with 2 different mutations), embedded in OCT and sectioned. The sections were trypsin digested and subjected to mass spectrometry imaging using a targeted approach to detect TGFBIp. MALDI-MSI identified peptides from TGFBIp that co-localized with the amyloid corneal deposits. In addition to the relative abundance data, the specific location of the peptides across the corneal sections as molecular signatures was also identified. Spatial distribution and intensity of the TGFBIp peptides showed differences between diseased and control models but also between the two LCD phenotypes. The TGFBIp peptide with <i>m</i>/<i>z</i> of 787.474 and <i>m</i>/<i>z</i> of 1179.579 showed increased expression in both LCD mutants compared to the controls. The peptide with <i>m</i>/<i>z</i> of 929.5 showed increased expression in the LCD phenotype with H626R mutation while the peptide with <i>m</i>/<i>z</i> of 1315.802 was abundant in the sample with R124C mutation. This initial report of 2D spatial protein signature and localization of TGFBIp may be expanded to other mutations to understand the proteolytic patterns of TGFBIp in different mutations. |
topic |
amyloid fibrils cornea lattice corneal dystrophy matrix-assisted laser desorption ionization (MALDI) mass spectrometry imaging (MSI) TGFBIp |
url |
https://www.mdpi.com/2297-8739/8/7/97 |
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