In vitro cross-linking of elastin peptides and molecular characterization of the resultant biomaterials

Andrea Heinz; Christoph K H Ruttkies; Günther Jahreis; Christoph U Schräder; Kanin Wichapong; Wolfgang Sippl; Fred W Keeley; Reinhard H H Neubert; Christian E H Schmelzer

doi:10.1016/j.bbagen.2013.01.014

In vitro cross-linking of elastin peptides and molecular characterization of the resultant biomaterials

Andrea Heinz, Christoph K H Ruttkies, Günther Jahreis, Christoph U Schräder, Kanin Wichapong, Wolfgang Sippl, Fred W Keeley, Reinhard H H Neubert, Christian E H Schmelzer

20 Citationer (Scopus)

Abstract

BACKGROUND: Elastin is a vital protein and the major component of elastic fibers which provides resilience to many vertebrate tissues. Elastin's structure and function are influenced by extensive cross-linking, however, the cross-linking pattern is still unknown.

METHODS: Small peptides containing reactive allysine residues based on sequences of cross-linking domains of human elastin were incubated in vitro to form cross-links characteristic of mature elastin. The resultant insoluble polymeric biomaterials were studied by scanning electron microscopy. Both, the supernatants of the samples and the insoluble polymers, after digestion with pancreatic elastase or trypsin, were furthermore comprehensively characterized on the molecular level using MALDI-TOF/TOF mass spectrometry.

RESULTS: MS(2) data was used to develop the software PolyLinX, which is able to sequence not only linear and bifunctionally cross-linked peptides, but for the first time also tri- and tetrafunctionally cross-linked species. Thus, it was possible to identify intra- and intermolecular cross-links including allysine aldols, dehydrolysinonorleucines and dehydromerodesmosines. The formation of the tetrafunctional cross-link desmosine or isodesmosine was unexpected, however, could be confirmed by tandem mass spectrometry and molecular dynamics simulations.

CONCLUSIONS: The study demonstrated that it is possible to produce biopolymers containing polyfunctional cross-links characteristic of mature elastin from small elastin peptides. MALDI-TOF/TOF mass spectrometry and the newly developed software PolyLinX proved suitable for sequencing of native cross-links in proteolytic digests of elastin-like biomaterials.

GENERAL SIGNIFICANCE: The study provides important insight into the formation of native elastin cross-links and represents a considerable step towards the characterization of the complex cross-linking pattern of mature elastin.

Originalsprog	Engelsk
Tidsskrift	B B A - Reviews on Cancer
Vol/bind	1830
Udgave nummer	4
Sider (fra-til)	2994-3004
Antal sider	11
ISSN	0006-3002
DOI	https://doi.org/10.1016/j.bbagen.2013.01.014
Status	Udgivet - apr. 2013

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10.1016/j.bbagen.2013.01.014

Citationsformater

@article{39f5637b6979427cacebea488e25619d,

title = "In vitro cross-linking of elastin peptides and molecular characterization of the resultant biomaterials",

abstract = "BACKGROUND: Elastin is a vital protein and the major component of elastic fibers which provides resilience to many vertebrate tissues. Elastin's structure and function are influenced by extensive cross-linking, however, the cross-linking pattern is still unknown.METHODS: Small peptides containing reactive allysine residues based on sequences of cross-linking domains of human elastin were incubated in vitro to form cross-links characteristic of mature elastin. The resultant insoluble polymeric biomaterials were studied by scanning electron microscopy. Both, the supernatants of the samples and the insoluble polymers, after digestion with pancreatic elastase or trypsin, were furthermore comprehensively characterized on the molecular level using MALDI-TOF/TOF mass spectrometry.RESULTS: MS(2) data was used to develop the software PolyLinX, which is able to sequence not only linear and bifunctionally cross-linked peptides, but for the first time also tri- and tetrafunctionally cross-linked species. Thus, it was possible to identify intra- and intermolecular cross-links including allysine aldols, dehydrolysinonorleucines and dehydromerodesmosines. The formation of the tetrafunctional cross-link desmosine or isodesmosine was unexpected, however, could be confirmed by tandem mass spectrometry and molecular dynamics simulations.CONCLUSIONS: The study demonstrated that it is possible to produce biopolymers containing polyfunctional cross-links characteristic of mature elastin from small elastin peptides. MALDI-TOF/TOF mass spectrometry and the newly developed software PolyLinX proved suitable for sequencing of native cross-links in proteolytic digests of elastin-like biomaterials.GENERAL SIGNIFICANCE: The study provides important insight into the formation of native elastin cross-links and represents a considerable step towards the characterization of the complex cross-linking pattern of mature elastin.",

keywords = "Amino Acid Sequence, Elastin, Humans, Molecular Dynamics Simulation, Molecular Sequence Data, Protein Structure, Tertiary, Software, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Journal Article, Research Support, Non-U.S. Gov't",

author = "Andrea Heinz and Ruttkies, {Christoph K H} and G{\"u}nther Jahreis and Schr{\"a}der, {Christoph U} and Kanin Wichapong and Wolfgang Sippl and Keeley, {Fred W} and Neubert, {Reinhard H H} and Schmelzer, {Christian E H}",

year = "2013",

month = apr,

doi = "10.1016/j.bbagen.2013.01.014",

language = "English",

volume = "1830",

pages = "2994--3004",

journal = "B B A - Reviews on Cancer",

issn = "0006-3002",

publisher = "Elsevier",

number = "4",

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TY - JOUR

T1 - In vitro cross-linking of elastin peptides and molecular characterization of the resultant biomaterials

AU - Heinz, Andrea

AU - Ruttkies, Christoph K H

AU - Jahreis, Günther

AU - Schräder, Christoph U

AU - Wichapong, Kanin

AU - Sippl, Wolfgang

AU - Keeley, Fred W

AU - Neubert, Reinhard H H

AU - Schmelzer, Christian E H

PY - 2013/4

Y1 - 2013/4

N2 - BACKGROUND: Elastin is a vital protein and the major component of elastic fibers which provides resilience to many vertebrate tissues. Elastin's structure and function are influenced by extensive cross-linking, however, the cross-linking pattern is still unknown.METHODS: Small peptides containing reactive allysine residues based on sequences of cross-linking domains of human elastin were incubated in vitro to form cross-links characteristic of mature elastin. The resultant insoluble polymeric biomaterials were studied by scanning electron microscopy. Both, the supernatants of the samples and the insoluble polymers, after digestion with pancreatic elastase or trypsin, were furthermore comprehensively characterized on the molecular level using MALDI-TOF/TOF mass spectrometry.RESULTS: MS(2) data was used to develop the software PolyLinX, which is able to sequence not only linear and bifunctionally cross-linked peptides, but for the first time also tri- and tetrafunctionally cross-linked species. Thus, it was possible to identify intra- and intermolecular cross-links including allysine aldols, dehydrolysinonorleucines and dehydromerodesmosines. The formation of the tetrafunctional cross-link desmosine or isodesmosine was unexpected, however, could be confirmed by tandem mass spectrometry and molecular dynamics simulations.CONCLUSIONS: The study demonstrated that it is possible to produce biopolymers containing polyfunctional cross-links characteristic of mature elastin from small elastin peptides. MALDI-TOF/TOF mass spectrometry and the newly developed software PolyLinX proved suitable for sequencing of native cross-links in proteolytic digests of elastin-like biomaterials.GENERAL SIGNIFICANCE: The study provides important insight into the formation of native elastin cross-links and represents a considerable step towards the characterization of the complex cross-linking pattern of mature elastin.

AB - BACKGROUND: Elastin is a vital protein and the major component of elastic fibers which provides resilience to many vertebrate tissues. Elastin's structure and function are influenced by extensive cross-linking, however, the cross-linking pattern is still unknown.METHODS: Small peptides containing reactive allysine residues based on sequences of cross-linking domains of human elastin were incubated in vitro to form cross-links characteristic of mature elastin. The resultant insoluble polymeric biomaterials were studied by scanning electron microscopy. Both, the supernatants of the samples and the insoluble polymers, after digestion with pancreatic elastase or trypsin, were furthermore comprehensively characterized on the molecular level using MALDI-TOF/TOF mass spectrometry.RESULTS: MS(2) data was used to develop the software PolyLinX, which is able to sequence not only linear and bifunctionally cross-linked peptides, but for the first time also tri- and tetrafunctionally cross-linked species. Thus, it was possible to identify intra- and intermolecular cross-links including allysine aldols, dehydrolysinonorleucines and dehydromerodesmosines. The formation of the tetrafunctional cross-link desmosine or isodesmosine was unexpected, however, could be confirmed by tandem mass spectrometry and molecular dynamics simulations.CONCLUSIONS: The study demonstrated that it is possible to produce biopolymers containing polyfunctional cross-links characteristic of mature elastin from small elastin peptides. MALDI-TOF/TOF mass spectrometry and the newly developed software PolyLinX proved suitable for sequencing of native cross-links in proteolytic digests of elastin-like biomaterials.GENERAL SIGNIFICANCE: The study provides important insight into the formation of native elastin cross-links and represents a considerable step towards the characterization of the complex cross-linking pattern of mature elastin.

KW - Amino Acid Sequence

KW - Elastin

KW - Humans

KW - Molecular Dynamics Simulation

KW - Molecular Sequence Data

KW - Protein Structure, Tertiary

KW - Software

KW - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1016/j.bbagen.2013.01.014

DO - 10.1016/j.bbagen.2013.01.014

M3 - Journal article

C2 - 23375722

SN - 0006-3002

VL - 1830

SP - 2994

EP - 3004

JO - B B A - Reviews on Cancer

JF - B B A - Reviews on Cancer

IS - 4

ER -

In vitro cross-linking of elastin peptides and molecular characterization of the resultant biomaterials

Abstract

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