Periplakin interactions with lens intermediate and beaded filaments

Kyoung Hye Yoon, Paul G FitzGerald

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

PURPOSE. The lens assembles two systems of intermediated filaments - vimentin intermediate filament (IF) and highly divergent, lens-specific beaded filament (BF) - sequentially as epithelial cells differentiate into fiber cells. The goal of this study was to identify linker proteins that integrate the different lens IF into the biology of the lens fiber cells. METHODS. Antibodies to periplakin were used in coimmunoprecipitation studies to identify proteins that complex with BF and IF in detergent extracts of mouse lens. GST-periplakin fusion proteins were used to confirm coimmunoprecipitation results. Yeast two-hybrid analysis was used to establish direct linkage between periplakin and BF/IF proteins and to narrow down binding domains. Immunocytochemistry was used to establish spatial and temporal coexpression of periplakin and BF/IF. RESULTS. Periplakin is found complexed to BF and IF in the lens. The COOH terminus of periplakin was shown to have a strong affinity for the CP49 rod 2 domain but not its head or rod 1 domains. Low-level affinity was seen between the filensin rod domain and periplakin. Periplakin localization in lens overlapped with BF and IF. CONCLUSIONS. Despite divergence in primary sequence, predicted secondary structure, and filament structure, CP49 has conserved the capacity to bind a common IF linker protein, periplakin, and shares that binding capacity with the other major lens IF protein, vimentin. This suggests that mutations in periplakin have the potential to emulate the cataract seen in lenses with defective BF proteins.

Original languageEnglish (US)
Pages (from-to)1283-1289
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Volume50
Issue number3
DOIs
StatePublished - Mar 2009

Fingerprint

Intermediate Filaments
Lenses
Intermediate Filament Proteins
Vimentin
Proteins
Detergents
Cataract
Yeasts
Epithelial Cells
Immunohistochemistry
Head
Mutation
Antibodies

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Periplakin interactions with lens intermediate and beaded filaments. / Yoon, Kyoung Hye; FitzGerald, Paul G.

In: Investigative Ophthalmology and Visual Science, Vol. 50, No. 3, 03.2009, p. 1283-1289.

Research output: Contribution to journalArticle

@article{e4684a56cd024695bfb758f44638a343,
title = "Periplakin interactions with lens intermediate and beaded filaments",
abstract = "PURPOSE. The lens assembles two systems of intermediated filaments - vimentin intermediate filament (IF) and highly divergent, lens-specific beaded filament (BF) - sequentially as epithelial cells differentiate into fiber cells. The goal of this study was to identify linker proteins that integrate the different lens IF into the biology of the lens fiber cells. METHODS. Antibodies to periplakin were used in coimmunoprecipitation studies to identify proteins that complex with BF and IF in detergent extracts of mouse lens. GST-periplakin fusion proteins were used to confirm coimmunoprecipitation results. Yeast two-hybrid analysis was used to establish direct linkage between periplakin and BF/IF proteins and to narrow down binding domains. Immunocytochemistry was used to establish spatial and temporal coexpression of periplakin and BF/IF. RESULTS. Periplakin is found complexed to BF and IF in the lens. The COOH terminus of periplakin was shown to have a strong affinity for the CP49 rod 2 domain but not its head or rod 1 domains. Low-level affinity was seen between the filensin rod domain and periplakin. Periplakin localization in lens overlapped with BF and IF. CONCLUSIONS. Despite divergence in primary sequence, predicted secondary structure, and filament structure, CP49 has conserved the capacity to bind a common IF linker protein, periplakin, and shares that binding capacity with the other major lens IF protein, vimentin. This suggests that mutations in periplakin have the potential to emulate the cataract seen in lenses with defective BF proteins.",
author = "Yoon, {Kyoung Hye} and FitzGerald, {Paul G}",
year = "2009",
month = "3",
doi = "10.1167/iovs.08-2894",
language = "English (US)",
volume = "50",
pages = "1283--1289",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "3",

}

TY - JOUR

T1 - Periplakin interactions with lens intermediate and beaded filaments

AU - Yoon, Kyoung Hye

AU - FitzGerald, Paul G

PY - 2009/3

Y1 - 2009/3

N2 - PURPOSE. The lens assembles two systems of intermediated filaments - vimentin intermediate filament (IF) and highly divergent, lens-specific beaded filament (BF) - sequentially as epithelial cells differentiate into fiber cells. The goal of this study was to identify linker proteins that integrate the different lens IF into the biology of the lens fiber cells. METHODS. Antibodies to periplakin were used in coimmunoprecipitation studies to identify proteins that complex with BF and IF in detergent extracts of mouse lens. GST-periplakin fusion proteins were used to confirm coimmunoprecipitation results. Yeast two-hybrid analysis was used to establish direct linkage between periplakin and BF/IF proteins and to narrow down binding domains. Immunocytochemistry was used to establish spatial and temporal coexpression of periplakin and BF/IF. RESULTS. Periplakin is found complexed to BF and IF in the lens. The COOH terminus of periplakin was shown to have a strong affinity for the CP49 rod 2 domain but not its head or rod 1 domains. Low-level affinity was seen between the filensin rod domain and periplakin. Periplakin localization in lens overlapped with BF and IF. CONCLUSIONS. Despite divergence in primary sequence, predicted secondary structure, and filament structure, CP49 has conserved the capacity to bind a common IF linker protein, periplakin, and shares that binding capacity with the other major lens IF protein, vimentin. This suggests that mutations in periplakin have the potential to emulate the cataract seen in lenses with defective BF proteins.

AB - PURPOSE. The lens assembles two systems of intermediated filaments - vimentin intermediate filament (IF) and highly divergent, lens-specific beaded filament (BF) - sequentially as epithelial cells differentiate into fiber cells. The goal of this study was to identify linker proteins that integrate the different lens IF into the biology of the lens fiber cells. METHODS. Antibodies to periplakin were used in coimmunoprecipitation studies to identify proteins that complex with BF and IF in detergent extracts of mouse lens. GST-periplakin fusion proteins were used to confirm coimmunoprecipitation results. Yeast two-hybrid analysis was used to establish direct linkage between periplakin and BF/IF proteins and to narrow down binding domains. Immunocytochemistry was used to establish spatial and temporal coexpression of periplakin and BF/IF. RESULTS. Periplakin is found complexed to BF and IF in the lens. The COOH terminus of periplakin was shown to have a strong affinity for the CP49 rod 2 domain but not its head or rod 1 domains. Low-level affinity was seen between the filensin rod domain and periplakin. Periplakin localization in lens overlapped with BF and IF. CONCLUSIONS. Despite divergence in primary sequence, predicted secondary structure, and filament structure, CP49 has conserved the capacity to bind a common IF linker protein, periplakin, and shares that binding capacity with the other major lens IF protein, vimentin. This suggests that mutations in periplakin have the potential to emulate the cataract seen in lenses with defective BF proteins.

UR - http://www.scopus.com/inward/record.url?scp=62649133355&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=62649133355&partnerID=8YFLogxK

U2 - 10.1167/iovs.08-2894

DO - 10.1167/iovs.08-2894

M3 - Article

VL - 50

SP - 1283

EP - 1289

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 3

ER -