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Esben Skipper Sørensen

Identification of transglutaminase reactive residues in human osteopontin and their role in polymerization

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Identification of transglutaminase reactive residues in human osteopontin and their role in polymerization. / Christensen, Brian ; Zachariae, Elias D; Scavenius, Carsten; Thybo, Morten; Callesen, Morten M; Kløverpris, Søren; Oxvig, Claus; Enghild, Jan Johannes; Sørensen, Esben Skipper.

In: PLOS ONE, Vol. 9, No. 11, e113650, 11.2014.

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@article{8c8d272c7e254cf3af4b190a49b9d79d,
title = "Identification of transglutaminase reactive residues in human osteopontin and their role in polymerization",
abstract = "Osteopontin (OPN) is a highly posttranslationally modified protein present in several tissues where it is implicated in numerous physiological processes. OPN primarily exerts its functions through interaction with integrins via the Arg-Gly-Asp and Ser-Val-Val-Tyr-Gly-Leu-Arg sequences located in the N-terminal part of the protein. OPN can be polymerized by the cross-linking enzyme transglutaminase 2 (TG2), and polymerization has been shown to enhance the biological activity of OPN. However, little is known about the reactivity and location of the glutamine and lysine residues involved in the TG2-mediated modification of OPN. Here we show that TG2 catalyses the incorporation of 5-(Biotinamido)pentylamine at glutamines in both the N- and C-terminal parts of OPN, whereas TG2 primarily incorporated the glutamine-donor peptide biotinyl-TVQQEL-OH into the C-terminal part of OPN. By mass spectrometric analyses we identified Gln34, Gln42, Gln193 and Gln248 as the major TG2 reactive glutamines in OPN. The distribution of reactive Gln and Lys residues in OPN proved to be important, as the full-length protein but not the physiologically highly active integrin-binding N-terminal part of OPN were able to polymerize in a TG2-mediated reaction. Collectively, these data provide important new molecular knowledge about the mechanism of OPN polymerization.",
author = "Brian Christensen and Zachariae, {Elias D} and Carsten Scavenius and Morten Thybo and Callesen, {Morten M} and S{\o}ren Kl{\o}verpris and Claus Oxvig and Enghild, {Jan Johannes} and S{\o}rensen, {Esben Skipper}",
year = "2014",
month = "11",
doi = "10.1371/journal.pone.0113650",
language = "English",
volume = "9",
journal = "P L o S One",
issn = "1932-6203",
publisher = "public library of science",
number = "11",

}

RIS

TY - JOUR

T1 - Identification of transglutaminase reactive residues in human osteopontin and their role in polymerization

AU - Christensen, Brian

AU - Zachariae, Elias D

AU - Scavenius, Carsten

AU - Thybo, Morten

AU - Callesen, Morten M

AU - Kløverpris, Søren

AU - Oxvig, Claus

AU - Enghild, Jan Johannes

AU - Sørensen, Esben Skipper

PY - 2014/11

Y1 - 2014/11

N2 - Osteopontin (OPN) is a highly posttranslationally modified protein present in several tissues where it is implicated in numerous physiological processes. OPN primarily exerts its functions through interaction with integrins via the Arg-Gly-Asp and Ser-Val-Val-Tyr-Gly-Leu-Arg sequences located in the N-terminal part of the protein. OPN can be polymerized by the cross-linking enzyme transglutaminase 2 (TG2), and polymerization has been shown to enhance the biological activity of OPN. However, little is known about the reactivity and location of the glutamine and lysine residues involved in the TG2-mediated modification of OPN. Here we show that TG2 catalyses the incorporation of 5-(Biotinamido)pentylamine at glutamines in both the N- and C-terminal parts of OPN, whereas TG2 primarily incorporated the glutamine-donor peptide biotinyl-TVQQEL-OH into the C-terminal part of OPN. By mass spectrometric analyses we identified Gln34, Gln42, Gln193 and Gln248 as the major TG2 reactive glutamines in OPN. The distribution of reactive Gln and Lys residues in OPN proved to be important, as the full-length protein but not the physiologically highly active integrin-binding N-terminal part of OPN were able to polymerize in a TG2-mediated reaction. Collectively, these data provide important new molecular knowledge about the mechanism of OPN polymerization.

AB - Osteopontin (OPN) is a highly posttranslationally modified protein present in several tissues where it is implicated in numerous physiological processes. OPN primarily exerts its functions through interaction with integrins via the Arg-Gly-Asp and Ser-Val-Val-Tyr-Gly-Leu-Arg sequences located in the N-terminal part of the protein. OPN can be polymerized by the cross-linking enzyme transglutaminase 2 (TG2), and polymerization has been shown to enhance the biological activity of OPN. However, little is known about the reactivity and location of the glutamine and lysine residues involved in the TG2-mediated modification of OPN. Here we show that TG2 catalyses the incorporation of 5-(Biotinamido)pentylamine at glutamines in both the N- and C-terminal parts of OPN, whereas TG2 primarily incorporated the glutamine-donor peptide biotinyl-TVQQEL-OH into the C-terminal part of OPN. By mass spectrometric analyses we identified Gln34, Gln42, Gln193 and Gln248 as the major TG2 reactive glutamines in OPN. The distribution of reactive Gln and Lys residues in OPN proved to be important, as the full-length protein but not the physiologically highly active integrin-binding N-terminal part of OPN were able to polymerize in a TG2-mediated reaction. Collectively, these data provide important new molecular knowledge about the mechanism of OPN polymerization.

U2 - 10.1371/journal.pone.0113650

DO - 10.1371/journal.pone.0113650

M3 - Journal article

VL - 9

JO - P L o S One

JF - P L o S One

SN - 1932-6203

IS - 11

M1 - e113650

ER -