The serine protease HtrA1 cleaves misfolded transforming growth factor β-induced protein (TGFBIp) and induces amyloid formation

Ebbe Toftgaard Poulsen; Nadia Sukusu Nielsen; Carsten Scavenius; Emilie Hage Mogensen; Michael W. Risør; Kasper Runager; Marie V. Lukassen; Casper B. Rasmussen; Gunna Christiansen; Mette Richner; Henrik Vorum; Jan J. Enghild

doi:10.1074/jbc.RA119.009050

The serine protease HtrA1 cleaves misfolded transforming growth factor β-induced protein (TGFBIp) and induces amyloid formation

Ebbe Toftgaard Poulsen, Nadia Sukusu Nielsen, Carsten Scavenius, Emilie Hage Mogensen, Michael W. Risør, Kasper Runager, Marie V. Lukassen, Casper B. Rasmussen, Gunna Christiansen, Mette Richner, Henrik Vorum, Jan J. Enghild^*

^*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

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Abstract

The serine protease high-temperature requirement protein A1 (HtrA1) is associated with protein-misfolding disorders such as Alzheimer's disease and transforming growth factor β-induced protein (TGFBIp)-linked corneal dystrophy. In this study, using several biochemical and biophysical approaches, including recombinant protein expression, LC-MS/MS and 2DE analyses, and thioflavin T (ThT) fluorescence assays for amyloid fibril detection, and FTIR assays, we investigated the role of HtrA1 both in normal TGFBIp turnover and in corneal amyloid formation. We show that HtrA1 can cleave WT TGFBIp but prefers amyloidogenic variants. Corneal TGFBIp is extensively processed in healthy people, resulting in C-terminal degradation products spanning the FAS1-4 domain of TGFBIp. We show here that HtrA1 cleaves the WT FAS1-4 domain only inefficiently, whereas the amyloidogenic FAS1-4 mutations transform this domain into a considerably better HTRA1 substrate. Moreover, HtrA1 cleavage of the mutant FAS1-4 domains generated peptides capable of forming in vitro amyloid aggregates. Significantly, these peptides have been previously identified in amyloid deposits in vivo, supporting the idea that HtrA1 is a causative agent for TGFBIp-associated amyloidosis in corneal dystrophy. In summary, our results indicate that TGFBIp is an HtrA1 substrate and that some mutations in the gene encoding TGFBIp cause aberrant HtrA1-mediated processing that results in amyloidogenesis in corneal dystrophies.

Original language	English
Journal	Journal of Biological Chemistry
Volume	294
Issue	31
Pages (from-to)	11817-11828
Number of pages	12
ISSN	0021-9258
DOIs	https://doi.org/10.1074/jbc.RA119.009050
Publication status	Published - Aug 2019

Keywords

BETA-IG-H3
BIGH3
CORNEA
HTRA1
IDENTIFICATION
LATTICE
MATRIX
MECHANISMS
MUTATIONS
PROTEOMICS
REVEALS
SUSCEPTIBILITY
amyloid
cornea
corneal dystrophy
eye disorder
granular corneal dystrophy (GCD)
high-temperature requirement protein A1 (HtrA1)
lattice corneal dystrophy (LCD)
protease
protein folding
protein misfolding
protein turnover
proteolytic processing
transforming growth factor beta-induced protein (TGFBIp)
Extracellular Matrix Proteins/chemistry
Humans
Chromatography, High Pressure Liquid
Amyloid/metabolism
Tandem Mass Spectrometry
High-Temperature Requirement A Serine Peptidase 1/genetics
Peptides/analysis
Aged, 80 and over
Protein Domains
Mutagenesis, Site-Directed
Recombinant Proteins/biosynthesis
Protein Folding
Corneal Diseases/metabolism
Transforming Growth Factor beta/chemistry
Cornea/metabolism

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10.1074/jbc.RA119.009050Licence: CC BY

The serine protease HtrA1 cleaves misfolded transforming growth factor β-induced protein (TGFBIp) and induces amyloid formationFinal published version, 2.86 MBLicence: CC BY

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Poulsen, E. T., Nielsen, N. S., Scavenius, C., Mogensen, E. H., Risør, M. W., Runager, K., Lukassen, M. V., Rasmussen, C. B., Christiansen, G., Richner, M., Vorum, H., & Enghild, J. J. (2019). The serine protease HtrA1 cleaves misfolded transforming growth factor β-induced protein (TGFBIp) and induces amyloid formation. Journal of Biological Chemistry, 294(31), 11817-11828. https://doi.org/10.1074/jbc.RA119.009050

@article{27ef2f4edc2b4d5c80414b65088443b9,

title = "The serine protease HtrA1 cleaves misfolded transforming growth factor β-induced protein (TGFBIp) and induces amyloid formation",

abstract = "The serine protease high-temperature requirement protein A1 (HtrA1) is associated with protein-misfolding disorders such as Alzheimer's disease and transforming growth factor β-induced protein (TGFBIp)-linked corneal dystrophy. In this study, using several biochemical and biophysical approaches, including recombinant protein expression, LC-MS/MS and 2DE analyses, and thioflavin T (ThT) fluorescence assays for amyloid fibril detection, and FTIR assays, we investigated the role of HtrA1 both in normal TGFBIp turnover and in corneal amyloid formation. We show that HtrA1 can cleave WT TGFBIp but prefers amyloidogenic variants. Corneal TGFBIp is extensively processed in healthy people, resulting in C-terminal degradation products spanning the FAS1-4 domain of TGFBIp. We show here that HtrA1 cleaves the WT FAS1-4 domain only inefficiently, whereas the amyloidogenic FAS1-4 mutations transform this domain into a considerably better HTRA1 substrate. Moreover, HtrA1 cleavage of the mutant FAS1-4 domains generated peptides capable of forming in vitro amyloid aggregates. Significantly, these peptides have been previously identified in amyloid deposits in vivo, supporting the idea that HtrA1 is a causative agent for TGFBIp-associated amyloidosis in corneal dystrophy. In summary, our results indicate that TGFBIp is an HtrA1 substrate and that some mutations in the gene encoding TGFBIp cause aberrant HtrA1-mediated processing that results in amyloidogenesis in corneal dystrophies. ",

keywords = "BETA-IG-H3, BIGH3, CORNEA, HTRA1, IDENTIFICATION, LATTICE, MATRIX, MECHANISMS, MUTATIONS, PROTEOMICS, REVEALS, SUSCEPTIBILITY, amyloid, cornea, corneal dystrophy, eye disorder, granular corneal dystrophy (GCD), high-temperature requirement protein A1 (HtrA1), lattice corneal dystrophy (LCD), protease, protein folding, protein misfolding, protein turnover, proteolytic processing, transforming growth factor beta-induced protein (TGFBIp), Extracellular Matrix Proteins/chemistry, Humans, Chromatography, High Pressure Liquid, Amyloid/metabolism, Tandem Mass Spectrometry, High-Temperature Requirement A Serine Peptidase 1/genetics, Peptides/analysis, Aged, 80 and over, Protein Domains, Mutagenesis, Site-Directed, Recombinant Proteins/biosynthesis, Protein Folding, Corneal Diseases/metabolism, Transforming Growth Factor beta/chemistry, Cornea/metabolism",

author = "Poulsen, {Ebbe Toftgaard} and Nielsen, {Nadia Sukusu} and Carsten Scavenius and Mogensen, {Emilie Hage} and Ris{\o}r, {Michael W.} and Kasper Runager and Lukassen, {Marie V.} and Rasmussen, {Casper B.} and Gunna Christiansen and Mette Richner and Henrik Vorum and Enghild, {Jan J.}",

note = "{\textcopyright} 2019 Poulsen et al.",

year = "2019",

month = aug,

doi = "10.1074/jbc.RA119.009050",

language = "English",

volume = "294",

pages = "11817--11828",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

publisher = "American Society for Biochemistry and Molecular Biology Inc.",

number = "31",

}

Poulsen, ET, Nielsen, NS , Scavenius, C, Mogensen, EH, Risør, MW, Runager, K, Lukassen, MV, Rasmussen, CB, Christiansen, G, Richner, M, Vorum, H & Enghild, JJ 2019, 'The serine protease HtrA1 cleaves misfolded transforming growth factor β-induced protein (TGFBIp) and induces amyloid formation', Journal of Biological Chemistry, vol. 294, no. 31, pp. 11817-11828. https://doi.org/10.1074/jbc.RA119.009050

The serine protease HtrA1 cleaves misfolded transforming growth factor β-induced protein (TGFBIp) and induces amyloid formation. / Poulsen, Ebbe Toftgaard; Nielsen, Nadia Sukusu ; Scavenius, Carsten et al.
In: Journal of Biological Chemistry, Vol. 294, No. 31, 08.2019, p. 11817-11828.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - The serine protease HtrA1 cleaves misfolded transforming growth factor β-induced protein (TGFBIp) and induces amyloid formation

AU - Poulsen, Ebbe Toftgaard

AU - Nielsen, Nadia Sukusu

AU - Scavenius, Carsten

AU - Mogensen, Emilie Hage

AU - Risør, Michael W.

AU - Runager, Kasper

AU - Lukassen, Marie V.

AU - Rasmussen, Casper B.

AU - Christiansen, Gunna

AU - Richner, Mette

AU - Vorum, Henrik

AU - Enghild, Jan J.

PY - 2019/8

Y1 - 2019/8

N2 - The serine protease high-temperature requirement protein A1 (HtrA1) is associated with protein-misfolding disorders such as Alzheimer's disease and transforming growth factor β-induced protein (TGFBIp)-linked corneal dystrophy. In this study, using several biochemical and biophysical approaches, including recombinant protein expression, LC-MS/MS and 2DE analyses, and thioflavin T (ThT) fluorescence assays for amyloid fibril detection, and FTIR assays, we investigated the role of HtrA1 both in normal TGFBIp turnover and in corneal amyloid formation. We show that HtrA1 can cleave WT TGFBIp but prefers amyloidogenic variants. Corneal TGFBIp is extensively processed in healthy people, resulting in C-terminal degradation products spanning the FAS1-4 domain of TGFBIp. We show here that HtrA1 cleaves the WT FAS1-4 domain only inefficiently, whereas the amyloidogenic FAS1-4 mutations transform this domain into a considerably better HTRA1 substrate. Moreover, HtrA1 cleavage of the mutant FAS1-4 domains generated peptides capable of forming in vitro amyloid aggregates. Significantly, these peptides have been previously identified in amyloid deposits in vivo, supporting the idea that HtrA1 is a causative agent for TGFBIp-associated amyloidosis in corneal dystrophy. In summary, our results indicate that TGFBIp is an HtrA1 substrate and that some mutations in the gene encoding TGFBIp cause aberrant HtrA1-mediated processing that results in amyloidogenesis in corneal dystrophies.

AB - The serine protease high-temperature requirement protein A1 (HtrA1) is associated with protein-misfolding disorders such as Alzheimer's disease and transforming growth factor β-induced protein (TGFBIp)-linked corneal dystrophy. In this study, using several biochemical and biophysical approaches, including recombinant protein expression, LC-MS/MS and 2DE analyses, and thioflavin T (ThT) fluorescence assays for amyloid fibril detection, and FTIR assays, we investigated the role of HtrA1 both in normal TGFBIp turnover and in corneal amyloid formation. We show that HtrA1 can cleave WT TGFBIp but prefers amyloidogenic variants. Corneal TGFBIp is extensively processed in healthy people, resulting in C-terminal degradation products spanning the FAS1-4 domain of TGFBIp. We show here that HtrA1 cleaves the WT FAS1-4 domain only inefficiently, whereas the amyloidogenic FAS1-4 mutations transform this domain into a considerably better HTRA1 substrate. Moreover, HtrA1 cleavage of the mutant FAS1-4 domains generated peptides capable of forming in vitro amyloid aggregates. Significantly, these peptides have been previously identified in amyloid deposits in vivo, supporting the idea that HtrA1 is a causative agent for TGFBIp-associated amyloidosis in corneal dystrophy. In summary, our results indicate that TGFBIp is an HtrA1 substrate and that some mutations in the gene encoding TGFBIp cause aberrant HtrA1-mediated processing that results in amyloidogenesis in corneal dystrophies.

KW - BETA-IG-H3

KW - BIGH3

KW - CORNEA

KW - HTRA1

KW - IDENTIFICATION

KW - LATTICE

KW - MATRIX

KW - MECHANISMS

KW - MUTATIONS

KW - PROTEOMICS

KW - REVEALS

KW - SUSCEPTIBILITY

KW - amyloid

KW - cornea

KW - corneal dystrophy

KW - eye disorder

KW - granular corneal dystrophy (GCD)

KW - high-temperature requirement protein A1 (HtrA1)

KW - lattice corneal dystrophy (LCD)

KW - protease

KW - protein folding

KW - protein misfolding

KW - protein turnover

KW - proteolytic processing

KW - transforming growth factor beta-induced protein (TGFBIp)

KW - Extracellular Matrix Proteins/chemistry

KW - Humans

KW - Chromatography, High Pressure Liquid

KW - Amyloid/metabolism

KW - Tandem Mass Spectrometry

KW - High-Temperature Requirement A Serine Peptidase 1/genetics

KW - Peptides/analysis

KW - Aged, 80 and over

KW - Protein Domains

KW - Mutagenesis, Site-Directed

KW - Recombinant Proteins/biosynthesis

KW - Protein Folding

KW - Corneal Diseases/metabolism

KW - Transforming Growth Factor beta/chemistry

KW - Cornea/metabolism

UR - http://www.scopus.com/inward/record.url?scp=85070745851&partnerID=8YFLogxK

U2 - 10.1074/jbc.RA119.009050

DO - 10.1074/jbc.RA119.009050

M3 - Journal article

C2 - 31197037

AN - SCOPUS:85070745851

SN - 0021-9258

VL - 294

SP - 11817

EP - 11828

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 31

ER -

The serine protease HtrA1 cleaves misfolded transforming growth factor β-induced protein (TGFBIp) and induces amyloid formation

Abstract

Keywords

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