In vitro cross-linking of elastin peptides and molecular characterization of the resultant biomaterials.
In vitro cross-linking of elastin peptides and molecular characterization of the resultant biomaterials.
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弹性蛋白肽的体外交联以及所得生物材料的分子表征
DOI：
10.1016/j.bbagen.2013.01.014
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
C.E.H. Schmelzer
中科院分区：
文献类型：
--
作者：
A. Heinz;C.K.H. Ruttkies;G. Jahreis;C.U. Schräder;K. Wichapong;W. Sippl;F.W. Keeley;R.H.H. Neubert;C.E.H. Schmelzer
弹力蛋白是一种重要的蛋白质，也是弹力纤维的主要成分，弹力纤维为许多脊椎动物组织提供弹性。弹性蛋白的结构和功能受到广泛交联的影响，但交联方式尚不清楚。方法根据人弹性蛋白的交联结构域序列，在体外培养含有活性丙氨酸残基的小肽，形成成熟弹性蛋白的交联特征。用扫描电镜对合成的不溶性高分子生物材料进行了研究。采用MALDI-TOF/TOF质谱法对样品的上清液和不溶性聚合物进行分子水平的全面表征。结果利用tsms2data开发了PolyLinX软件，该软件不仅能够对线性和双功能交联肽进行测序，而且首次能够对三功能交联和四功能交联物种进行测序。因此，可以鉴定分子内和分子间的交联，包括丙氨酸醛醇、脱氢氨基亮氨酸和脱氢氨基葡萄糖。四功能交联桥mosine或异桥mosine的形成是意料之外的，然而，可以通过串联质谱和分子动力学模拟来证实。结论利用弹性蛋白小肽制备具有成熟弹性蛋白多功能交联特性的生物聚合物是可行的。MALDI-TOF/TOF质谱法和新开发的软件PolyLinX被证明适用于弹性蛋白样生物材料蛋白水解酶的天然交联测序。该研究对天然弹性蛋白交联的形成提供了重要的见解，并代表了成熟弹性蛋白复杂交联模式表征的重要一步。
BACKGROUNDElastin 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.METHODSSmall 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.RESULTSMS2data 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.CONCLUSIONSThe 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 SIGNIFICANCEThe 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.
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DOI：
10.1016/0305-0491(81)90158-9
发表时间：
1981
期刊：
Comparative Biochemistry and Physiology B
影响因子：
--
作者：
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通讯作者：
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DOI：
10.1016/s0021-9258(18)82207-4
发表时间：
1993-05
期刊：
The Journal of biological chemistry
影响因子：
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作者：
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通讯作者：
D. Bedell-Hogan;P. Trackman;William AbramsS;Joel RosenbloomS;Herbert Kagang
影响因子：
5.4
作者：
Heinz, Andrea;Jung, Michael C.;Schmelzer, Christian E. H.
通讯作者：
Schmelzer, Christian E. H.
DOI：
10.1098/rstb.2001.1027
发表时间：
2002-02-28
影响因子：
6.3
作者：
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通讯作者：
Woodhouse, KA
影响因子：
48
作者：
O. Rinner;J. Seebacher;T. Walzthoeni;Lukas N. Mueller;Martin Beck;A. Schmidt;Markus Mueller;R. Aebersold
通讯作者：
O. Rinner;J. Seebacher;T. Walzthoeni;Lukas N. Mueller;Martin Beck;A. Schmidt;Markus Mueller;R. Aebersold