A novel function for transglutaminase 1: Attachment of long-chain ω- hydroxyceramides to involucrin by ester bond formation

Z. Nemes, L. N. Marekov, L. Fésüs, P. M. Steinert

Research output: Contribution to journalArticle

193 Citations (Scopus)

Abstract

Transglutaminases (TGases) are defined as enzymes capable of forming isopeptide bonds by transfer of an amine onto glutaminyl residues of a protein. Here we show that the membrane-bound form of the TGase 1 enzyme can also form ester bonds between specific glutaminyl residues of human involucrin and a synthetic analog of epidermal specific ω-hydroxyceramides. The formation of a ≃5-nm-thick lipid envelope on the surface of epidermal keratinocytes is an important component of normal barrier function. The lipid envelope consists of ω-hydroxyceramides covalently linked by ester bonds to cornified envelope proteins, most abundantly to involucrin. We synthesized an analog of natural ω-hydroxyceramides N-[16-(16- hydroxyhexadecyl)oxypalmitoyl]-sphingosine (lipid Z). When recombinant human TGase 1 and involucrin were reacted on the surface of synthetic lipid vesicles containing lipid Z, lipid Z was attached to involucrin and formed saponifiable protein-lipid adducts. By mass spectroscopy and sequencing of tryptic lipopeptides, the ester linkage formation used involucrin glutamine residues 107, 118, 122, 133, and 496 by converting the γ-carboxamido groups to lipid esters. Several of these residues have been found previously to be attached to ceramides in vivo. Mass spectrometric analysis after acetonide derivatization also revealed that ester formation involved primarily the ω- hydroxyl group of lipid Z. Our data reveal a dual role for TGase 1 in epidermal barrier formation and provide insights into the pathophysiology of lamellar ichthyosis resulting from defects of TGase 1 enzyme.

Original languageEnglish
Pages (from-to)8402-8407
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number15
DOIs
Publication statusPublished - Jul 20 1999

Fingerprint

Esters
Lipids
Enzymes
Lamellar Ichthyosis
Lipopeptides
Proteins
Sphingosine
Transglutaminases
Ceramides
Glutamine
Keratinocytes
Hydroxyl Radical
Amines
Mass Spectrometry
involucrin
transglutaminase 1
Membranes
lipid Z

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

A novel function for transglutaminase 1 : Attachment of long-chain ω- hydroxyceramides to involucrin by ester bond formation. / Nemes, Z.; Marekov, L. N.; Fésüs, L.; Steinert, P. M.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, No. 15, 20.07.1999, p. 8402-8407.

Research output: Contribution to journalArticle

@article{5cc66abf17dd4af79395bf3e7c91945f,
title = "A novel function for transglutaminase 1: Attachment of long-chain ω- hydroxyceramides to involucrin by ester bond formation",
abstract = "Transglutaminases (TGases) are defined as enzymes capable of forming isopeptide bonds by transfer of an amine onto glutaminyl residues of a protein. Here we show that the membrane-bound form of the TGase 1 enzyme can also form ester bonds between specific glutaminyl residues of human involucrin and a synthetic analog of epidermal specific ω-hydroxyceramides. The formation of a ≃5-nm-thick lipid envelope on the surface of epidermal keratinocytes is an important component of normal barrier function. The lipid envelope consists of ω-hydroxyceramides covalently linked by ester bonds to cornified envelope proteins, most abundantly to involucrin. We synthesized an analog of natural ω-hydroxyceramides N-[16-(16- hydroxyhexadecyl)oxypalmitoyl]-sphingosine (lipid Z). When recombinant human TGase 1 and involucrin were reacted on the surface of synthetic lipid vesicles containing lipid Z, lipid Z was attached to involucrin and formed saponifiable protein-lipid adducts. By mass spectroscopy and sequencing of tryptic lipopeptides, the ester linkage formation used involucrin glutamine residues 107, 118, 122, 133, and 496 by converting the γ-carboxamido groups to lipid esters. Several of these residues have been found previously to be attached to ceramides in vivo. Mass spectrometric analysis after acetonide derivatization also revealed that ester formation involved primarily the ω- hydroxyl group of lipid Z. Our data reveal a dual role for TGase 1 in epidermal barrier formation and provide insights into the pathophysiology of lamellar ichthyosis resulting from defects of TGase 1 enzyme.",
author = "Z. Nemes and Marekov, {L. N.} and L. F{\'e}s{\"u}s and Steinert, {P. M.}",
year = "1999",
month = "7",
day = "20",
doi = "10.1073/pnas.96.15.8402",
language = "English",
volume = "96",
pages = "8402--8407",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "15",

}

TY - JOUR

T1 - A novel function for transglutaminase 1

T2 - Attachment of long-chain ω- hydroxyceramides to involucrin by ester bond formation

AU - Nemes, Z.

AU - Marekov, L. N.

AU - Fésüs, L.

AU - Steinert, P. M.

PY - 1999/7/20

Y1 - 1999/7/20

N2 - Transglutaminases (TGases) are defined as enzymes capable of forming isopeptide bonds by transfer of an amine onto glutaminyl residues of a protein. Here we show that the membrane-bound form of the TGase 1 enzyme can also form ester bonds between specific glutaminyl residues of human involucrin and a synthetic analog of epidermal specific ω-hydroxyceramides. The formation of a ≃5-nm-thick lipid envelope on the surface of epidermal keratinocytes is an important component of normal barrier function. The lipid envelope consists of ω-hydroxyceramides covalently linked by ester bonds to cornified envelope proteins, most abundantly to involucrin. We synthesized an analog of natural ω-hydroxyceramides N-[16-(16- hydroxyhexadecyl)oxypalmitoyl]-sphingosine (lipid Z). When recombinant human TGase 1 and involucrin were reacted on the surface of synthetic lipid vesicles containing lipid Z, lipid Z was attached to involucrin and formed saponifiable protein-lipid adducts. By mass spectroscopy and sequencing of tryptic lipopeptides, the ester linkage formation used involucrin glutamine residues 107, 118, 122, 133, and 496 by converting the γ-carboxamido groups to lipid esters. Several of these residues have been found previously to be attached to ceramides in vivo. Mass spectrometric analysis after acetonide derivatization also revealed that ester formation involved primarily the ω- hydroxyl group of lipid Z. Our data reveal a dual role for TGase 1 in epidermal barrier formation and provide insights into the pathophysiology of lamellar ichthyosis resulting from defects of TGase 1 enzyme.

AB - Transglutaminases (TGases) are defined as enzymes capable of forming isopeptide bonds by transfer of an amine onto glutaminyl residues of a protein. Here we show that the membrane-bound form of the TGase 1 enzyme can also form ester bonds between specific glutaminyl residues of human involucrin and a synthetic analog of epidermal specific ω-hydroxyceramides. The formation of a ≃5-nm-thick lipid envelope on the surface of epidermal keratinocytes is an important component of normal barrier function. The lipid envelope consists of ω-hydroxyceramides covalently linked by ester bonds to cornified envelope proteins, most abundantly to involucrin. We synthesized an analog of natural ω-hydroxyceramides N-[16-(16- hydroxyhexadecyl)oxypalmitoyl]-sphingosine (lipid Z). When recombinant human TGase 1 and involucrin were reacted on the surface of synthetic lipid vesicles containing lipid Z, lipid Z was attached to involucrin and formed saponifiable protein-lipid adducts. By mass spectroscopy and sequencing of tryptic lipopeptides, the ester linkage formation used involucrin glutamine residues 107, 118, 122, 133, and 496 by converting the γ-carboxamido groups to lipid esters. Several of these residues have been found previously to be attached to ceramides in vivo. Mass spectrometric analysis after acetonide derivatization also revealed that ester formation involved primarily the ω- hydroxyl group of lipid Z. Our data reveal a dual role for TGase 1 in epidermal barrier formation and provide insights into the pathophysiology of lamellar ichthyosis resulting from defects of TGase 1 enzyme.

UR - http://www.scopus.com/inward/record.url?scp=0033587684&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033587684&partnerID=8YFLogxK

U2 - 10.1073/pnas.96.15.8402

DO - 10.1073/pnas.96.15.8402

M3 - Article

C2 - 10411887

AN - SCOPUS:0033587684

VL - 96

SP - 8402

EP - 8407

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 15

ER -