Liver disease-associated keratin 8 and 18 mutations modulate keratin acetylation and methylation

Kwi Hoon Jang, Han Na Yoon, Jongeun Lee, Hayan Yi, Sang Yoon Park, So Young Lee, Younglan Lim, Hyoung Joo Lee, Jin Won Cho, Young-Ki Paik, Williams S. Hancock, Nam-on Ku

Research output: Contribution to journalArticle

Abstract

Keratin 8 (K8) and keratin 18 (K18) are the intermediate filament proteins whose phosphorylation/transamidation associate with their aggregation in Mallory-Denk bodies found in patients with various liver diseases. However, the functions of other post-translational modifications in keratins related to liver diseases have not been fully elucidated. Here, using a site-specific mutation assay combined with nano-liquid chromatography-tandem mass spectrometry, we identified K8-Lys108 and K18-Lys187/426 as acetylation sites, and K8-Arg47 and K18-Arg55 as methylation sites. Keratin mutation (Arg-to-Lys/Ala) at the methylation sites, but not the acetylation sites, led to decreased stability of the keratin protein. We compared keratin acetylation/methylation in liver disease-associated keratin variants. The acetylation of K8 variants increased or decreased to various extents, whereas the methylation of K18-del65-72 and K18-I150V variants increased. Notably, the highly acetylated/methylated K18-I150V variant was less soluble and exhibited unusually prolonged protein stability, which suggests that additional acetylation of highly methylated keratins has a synergistic effect on prolonged stability. Therefore, the different levels of acetylation/methylation of the liver disease-associated variants regulate keratin protein stability. These findings extend our understanding of how disease-associated mutations in keratins modulate keratin acetylation and methylation, which may contribute to disease pathogenesis.-Jang, K.-H., Yoon, H.-N., Lee, J., Yi, H., Park, S.-Y., Lee, S.-Y., Lim, Y., Lee, H.-J., Cho, J.-W., Paik, Y.-K., Hancock, W. S., Ku, N.-O. Liver disease-associated keratin 8 and 18 mutations modulate keratin acetylation and methylation.

Original languageEnglish
Pages (from-to)9030-9043
Number of pages14
JournalFASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume33
Issue number8
DOIs
Publication statusPublished - 2019 Aug 1

Fingerprint

Keratin-8
Keratin-18
Acetylation
Methylation
Keratins
Liver
Liver Diseases
Mutation
Protein Stability
Mallory Bodies
Intermediate Filament Proteins
Phosphorylation
Proteins
Liquid chromatography
Post Translational Protein Processing
Tandem Mass Spectrometry
Liquid Chromatography
Mass spectrometry
Assays
Agglomeration

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Jang, Kwi Hoon ; Yoon, Han Na ; Lee, Jongeun ; Yi, Hayan ; Park, Sang Yoon ; Lee, So Young ; Lim, Younglan ; Lee, Hyoung Joo ; Cho, Jin Won ; Paik, Young-Ki ; Hancock, Williams S. ; Ku, Nam-on. / Liver disease-associated keratin 8 and 18 mutations modulate keratin acetylation and methylation. In: FASEB journal : official publication of the Federation of American Societies for Experimental Biology. 2019 ; Vol. 33, No. 8. pp. 9030-9043.
@article{a9c1aae8d1bf41b59e264ef8b49338a7,
title = "Liver disease-associated keratin 8 and 18 mutations modulate keratin acetylation and methylation",
abstract = "Keratin 8 (K8) and keratin 18 (K18) are the intermediate filament proteins whose phosphorylation/transamidation associate with their aggregation in Mallory-Denk bodies found in patients with various liver diseases. However, the functions of other post-translational modifications in keratins related to liver diseases have not been fully elucidated. Here, using a site-specific mutation assay combined with nano-liquid chromatography-tandem mass spectrometry, we identified K8-Lys108 and K18-Lys187/426 as acetylation sites, and K8-Arg47 and K18-Arg55 as methylation sites. Keratin mutation (Arg-to-Lys/Ala) at the methylation sites, but not the acetylation sites, led to decreased stability of the keratin protein. We compared keratin acetylation/methylation in liver disease-associated keratin variants. The acetylation of K8 variants increased or decreased to various extents, whereas the methylation of K18-del65-72 and K18-I150V variants increased. Notably, the highly acetylated/methylated K18-I150V variant was less soluble and exhibited unusually prolonged protein stability, which suggests that additional acetylation of highly methylated keratins has a synergistic effect on prolonged stability. Therefore, the different levels of acetylation/methylation of the liver disease-associated variants regulate keratin protein stability. These findings extend our understanding of how disease-associated mutations in keratins modulate keratin acetylation and methylation, which may contribute to disease pathogenesis.-Jang, K.-H., Yoon, H.-N., Lee, J., Yi, H., Park, S.-Y., Lee, S.-Y., Lim, Y., Lee, H.-J., Cho, J.-W., Paik, Y.-K., Hancock, W. S., Ku, N.-O. Liver disease-associated keratin 8 and 18 mutations modulate keratin acetylation and methylation.",
author = "Jang, {Kwi Hoon} and Yoon, {Han Na} and Jongeun Lee and Hayan Yi and Park, {Sang Yoon} and Lee, {So Young} and Younglan Lim and Lee, {Hyoung Joo} and Cho, {Jin Won} and Young-Ki Paik and Hancock, {Williams S.} and Nam-on Ku",
year = "2019",
month = "8",
day = "1",
doi = "10.1096/fj.201800263RR",
language = "English",
volume = "33",
pages = "9030--9043",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "8",

}

Liver disease-associated keratin 8 and 18 mutations modulate keratin acetylation and methylation. / Jang, Kwi Hoon; Yoon, Han Na; Lee, Jongeun; Yi, Hayan; Park, Sang Yoon; Lee, So Young; Lim, Younglan; Lee, Hyoung Joo; Cho, Jin Won; Paik, Young-Ki; Hancock, Williams S.; Ku, Nam-on.

In: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 33, No. 8, 01.08.2019, p. 9030-9043.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Liver disease-associated keratin 8 and 18 mutations modulate keratin acetylation and methylation

AU - Jang, Kwi Hoon

AU - Yoon, Han Na

AU - Lee, Jongeun

AU - Yi, Hayan

AU - Park, Sang Yoon

AU - Lee, So Young

AU - Lim, Younglan

AU - Lee, Hyoung Joo

AU - Cho, Jin Won

AU - Paik, Young-Ki

AU - Hancock, Williams S.

AU - Ku, Nam-on

PY - 2019/8/1

Y1 - 2019/8/1

N2 - Keratin 8 (K8) and keratin 18 (K18) are the intermediate filament proteins whose phosphorylation/transamidation associate with their aggregation in Mallory-Denk bodies found in patients with various liver diseases. However, the functions of other post-translational modifications in keratins related to liver diseases have not been fully elucidated. Here, using a site-specific mutation assay combined with nano-liquid chromatography-tandem mass spectrometry, we identified K8-Lys108 and K18-Lys187/426 as acetylation sites, and K8-Arg47 and K18-Arg55 as methylation sites. Keratin mutation (Arg-to-Lys/Ala) at the methylation sites, but not the acetylation sites, led to decreased stability of the keratin protein. We compared keratin acetylation/methylation in liver disease-associated keratin variants. The acetylation of K8 variants increased or decreased to various extents, whereas the methylation of K18-del65-72 and K18-I150V variants increased. Notably, the highly acetylated/methylated K18-I150V variant was less soluble and exhibited unusually prolonged protein stability, which suggests that additional acetylation of highly methylated keratins has a synergistic effect on prolonged stability. Therefore, the different levels of acetylation/methylation of the liver disease-associated variants regulate keratin protein stability. These findings extend our understanding of how disease-associated mutations in keratins modulate keratin acetylation and methylation, which may contribute to disease pathogenesis.-Jang, K.-H., Yoon, H.-N., Lee, J., Yi, H., Park, S.-Y., Lee, S.-Y., Lim, Y., Lee, H.-J., Cho, J.-W., Paik, Y.-K., Hancock, W. S., Ku, N.-O. Liver disease-associated keratin 8 and 18 mutations modulate keratin acetylation and methylation.

AB - Keratin 8 (K8) and keratin 18 (K18) are the intermediate filament proteins whose phosphorylation/transamidation associate with their aggregation in Mallory-Denk bodies found in patients with various liver diseases. However, the functions of other post-translational modifications in keratins related to liver diseases have not been fully elucidated. Here, using a site-specific mutation assay combined with nano-liquid chromatography-tandem mass spectrometry, we identified K8-Lys108 and K18-Lys187/426 as acetylation sites, and K8-Arg47 and K18-Arg55 as methylation sites. Keratin mutation (Arg-to-Lys/Ala) at the methylation sites, but not the acetylation sites, led to decreased stability of the keratin protein. We compared keratin acetylation/methylation in liver disease-associated keratin variants. The acetylation of K8 variants increased or decreased to various extents, whereas the methylation of K18-del65-72 and K18-I150V variants increased. Notably, the highly acetylated/methylated K18-I150V variant was less soluble and exhibited unusually prolonged protein stability, which suggests that additional acetylation of highly methylated keratins has a synergistic effect on prolonged stability. Therefore, the different levels of acetylation/methylation of the liver disease-associated variants regulate keratin protein stability. These findings extend our understanding of how disease-associated mutations in keratins modulate keratin acetylation and methylation, which may contribute to disease pathogenesis.-Jang, K.-H., Yoon, H.-N., Lee, J., Yi, H., Park, S.-Y., Lee, S.-Y., Lim, Y., Lee, H.-J., Cho, J.-W., Paik, Y.-K., Hancock, W. S., Ku, N.-O. Liver disease-associated keratin 8 and 18 mutations modulate keratin acetylation and methylation.

UR - http://www.scopus.com/inward/record.url?scp=85070787310&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85070787310&partnerID=8YFLogxK

U2 - 10.1096/fj.201800263RR

DO - 10.1096/fj.201800263RR

M3 - Article

C2 - 31199680

AN - SCOPUS:85070787310

VL - 33

SP - 9030

EP - 9043

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 8

ER -