Mutation in transforming growth factor beta induced protein associated with granular corneal dystrophy type 1 reduces the proteolytic susceptibility through local structural stabilization

Jarl Underhaug, Heidi Koldsø, Kasper Runager, Jakob Toudahl Nielsen, Charlotte S Sørensen, Torsten Kristensen, Daniel E Otzen, Henrik Karring, Anders Malmendal, Birgit Schiøtt, Jan J Enghild, Niels Chr Nielsen

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Resumé

Hereditary mutations in the transforming growth factor beta induced (TGFBI) gene cause phenotypically distinct corneal dystrophies characterized by protein deposition in cornea. We show here that the Arg555Trp mutant of the fourth fasciclin 1 (FAS1-4) domain of the protein (TGFBIp/keratoepithelin/βig-h3), associated with granular corneal dystrophy type 1, is significantly less susceptible to proteolysis by thermolysin and trypsin than the WT domain. High-resolution liquid-state NMR of the WT and Arg555Trp mutant FAS1-4 domains revealed very similar structures except for the region around position 555. The Arg555Trp substitution causes Trp555 to be buried in an otherwise empty hydrophobic cavity of the FAS1-4 domain. The first thermolysin cleavage in the core of the FAS1-4 domain occurs on the N-terminal side of Leu558 adjacent to the Arg555 mutation. MD simulations indicated that the C-terminal end of helix α3' containing this cleavage site is less flexible in the mutant domain, explaining the observed proteolytic resistance. This structural change also alters the electrostatic properties, which may explain increased propensity of the mutant to aggregate in vitro with 2,2,2-trifluoroethanol. Based on our results we propose that the Arg555Trp mutation disrupts the normal degradation/turnover of corneal TGFBIp, leading to accumulation and increased propensity to aggregate through electrostatic interactions.

OriginalsprogEngelsk
TidsskriftBBA General Subjects
Vol/bind1834
Udgave nummer12
Sider (fra-til)2812-22
Antal sider11
ISSN0304-4165
DOI
StatusUdgivet - dec. 2013
Udgivet eksterntJa

Citer dette

Underhaug, Jarl ; Koldsø, Heidi ; Runager, Kasper ; Nielsen, Jakob Toudahl ; Sørensen, Charlotte S ; Kristensen, Torsten ; Otzen, Daniel E ; Karring, Henrik ; Malmendal, Anders ; Schiøtt, Birgit ; Enghild, Jan J ; Nielsen, Niels Chr. / Mutation in transforming growth factor beta induced protein associated with granular corneal dystrophy type 1 reduces the proteolytic susceptibility through local structural stabilization. I: BBA General Subjects. 2013 ; Bind 1834, Nr. 12. s. 2812-22.
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title = "Mutation in transforming growth factor beta induced protein associated with granular corneal dystrophy type 1 reduces the proteolytic susceptibility through local structural stabilization",
abstract = "Hereditary mutations in the transforming growth factor beta induced (TGFBI) gene cause phenotypically distinct corneal dystrophies characterized by protein deposition in cornea. We show here that the Arg555Trp mutant of the fourth fasciclin 1 (FAS1-4) domain of the protein (TGFBIp/keratoepithelin/βig-h3), associated with granular corneal dystrophy type 1, is significantly less susceptible to proteolysis by thermolysin and trypsin than the WT domain. High-resolution liquid-state NMR of the WT and Arg555Trp mutant FAS1-4 domains revealed very similar structures except for the region around position 555. The Arg555Trp substitution causes Trp555 to be buried in an otherwise empty hydrophobic cavity of the FAS1-4 domain. The first thermolysin cleavage in the core of the FAS1-4 domain occurs on the N-terminal side of Leu558 adjacent to the Arg555 mutation. MD simulations indicated that the C-terminal end of helix α3' containing this cleavage site is less flexible in the mutant domain, explaining the observed proteolytic resistance. This structural change also alters the electrostatic properties, which may explain increased propensity of the mutant to aggregate in vitro with 2,2,2-trifluoroethanol. Based on our results we propose that the Arg555Trp mutation disrupts the normal degradation/turnover of corneal TGFBIp, leading to accumulation and increased propensity to aggregate through electrostatic interactions.",
author = "Jarl Underhaug and Heidi Kolds{\o} and Kasper Runager and Nielsen, {Jakob Toudahl} and S{\o}rensen, {Charlotte S} and Torsten Kristensen and Otzen, {Daniel E} and Henrik Karring and Anders Malmendal and Birgit Schi{\o}tt and Enghild, {Jan J} and Nielsen, {Niels Chr}",
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year = "2013",
month = "12",
doi = "10.1016/j.bbapap.2013.10.008",
language = "English",
volume = "1834",
pages = "2812--22",
journal = "B B A - General Subjects",
issn = "0304-4165",
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Underhaug, J, Koldsø, H, Runager, K, Nielsen, JT, Sørensen, CS, Kristensen, T, Otzen, DE, Karring, H, Malmendal, A, Schiøtt, B, Enghild, JJ & Nielsen, NC 2013, 'Mutation in transforming growth factor beta induced protein associated with granular corneal dystrophy type 1 reduces the proteolytic susceptibility through local structural stabilization', BBA General Subjects, bind 1834, nr. 12, s. 2812-22. https://doi.org/10.1016/j.bbapap.2013.10.008

Mutation in transforming growth factor beta induced protein associated with granular corneal dystrophy type 1 reduces the proteolytic susceptibility through local structural stabilization. / Underhaug, Jarl; Koldsø, Heidi; Runager, Kasper; Nielsen, Jakob Toudahl; Sørensen, Charlotte S; Kristensen, Torsten; Otzen, Daniel E; Karring, Henrik; Malmendal, Anders; Schiøtt, Birgit; Enghild, Jan J; Nielsen, Niels Chr.

I: BBA General Subjects, Bind 1834, Nr. 12, 12.2013, s. 2812-22.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Mutation in transforming growth factor beta induced protein associated with granular corneal dystrophy type 1 reduces the proteolytic susceptibility through local structural stabilization

AU - Underhaug, Jarl

AU - Koldsø, Heidi

AU - Runager, Kasper

AU - Nielsen, Jakob Toudahl

AU - Sørensen, Charlotte S

AU - Kristensen, Torsten

AU - Otzen, Daniel E

AU - Karring, Henrik

AU - Malmendal, Anders

AU - Schiøtt, Birgit

AU - Enghild, Jan J

AU - Nielsen, Niels Chr

N1 - © 2013.

PY - 2013/12

Y1 - 2013/12

N2 - Hereditary mutations in the transforming growth factor beta induced (TGFBI) gene cause phenotypically distinct corneal dystrophies characterized by protein deposition in cornea. We show here that the Arg555Trp mutant of the fourth fasciclin 1 (FAS1-4) domain of the protein (TGFBIp/keratoepithelin/βig-h3), associated with granular corneal dystrophy type 1, is significantly less susceptible to proteolysis by thermolysin and trypsin than the WT domain. High-resolution liquid-state NMR of the WT and Arg555Trp mutant FAS1-4 domains revealed very similar structures except for the region around position 555. The Arg555Trp substitution causes Trp555 to be buried in an otherwise empty hydrophobic cavity of the FAS1-4 domain. The first thermolysin cleavage in the core of the FAS1-4 domain occurs on the N-terminal side of Leu558 adjacent to the Arg555 mutation. MD simulations indicated that the C-terminal end of helix α3' containing this cleavage site is less flexible in the mutant domain, explaining the observed proteolytic resistance. This structural change also alters the electrostatic properties, which may explain increased propensity of the mutant to aggregate in vitro with 2,2,2-trifluoroethanol. Based on our results we propose that the Arg555Trp mutation disrupts the normal degradation/turnover of corneal TGFBIp, leading to accumulation and increased propensity to aggregate through electrostatic interactions.

AB - Hereditary mutations in the transforming growth factor beta induced (TGFBI) gene cause phenotypically distinct corneal dystrophies characterized by protein deposition in cornea. We show here that the Arg555Trp mutant of the fourth fasciclin 1 (FAS1-4) domain of the protein (TGFBIp/keratoepithelin/βig-h3), associated with granular corneal dystrophy type 1, is significantly less susceptible to proteolysis by thermolysin and trypsin than the WT domain. High-resolution liquid-state NMR of the WT and Arg555Trp mutant FAS1-4 domains revealed very similar structures except for the region around position 555. The Arg555Trp substitution causes Trp555 to be buried in an otherwise empty hydrophobic cavity of the FAS1-4 domain. The first thermolysin cleavage in the core of the FAS1-4 domain occurs on the N-terminal side of Leu558 adjacent to the Arg555 mutation. MD simulations indicated that the C-terminal end of helix α3' containing this cleavage site is less flexible in the mutant domain, explaining the observed proteolytic resistance. This structural change also alters the electrostatic properties, which may explain increased propensity of the mutant to aggregate in vitro with 2,2,2-trifluoroethanol. Based on our results we propose that the Arg555Trp mutation disrupts the normal degradation/turnover of corneal TGFBIp, leading to accumulation and increased propensity to aggregate through electrostatic interactions.

U2 - 10.1016/j.bbapap.2013.10.008

DO - 10.1016/j.bbapap.2013.10.008

M3 - Journal article

VL - 1834

SP - 2812

EP - 2822

JO - B B A - General Subjects

JF - B B A - General Subjects

SN - 0304-4165

IS - 12

ER -