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Title: Structural investigations on native collagen type I fibrils using AFM

Abstract

This study was carried out to determine the elastic properties of single collagen type I fibrils with the use of atomic force microscopy (AFM). Native collagen fibrils were formed by self-assembly in vitro characterized with the AFM. To confirm the inner assembly of the collagen fibrils, the AFM was used as a microdissection tool. Native collagen type I fibrils were dissected and the inner core uncovered. To determine the elastic properties of collagen fibrils the tip of the AFM was used as a nanoindentor by recording force-displacement curves. Measurements were done on the outer shell and in the core of the fibril. The structural investigations revealed the banding of the shell also in the core of native collagen fibrils. Nanoindentation experiments showed the same Young's modulus on the shell as well as in the core of the investigated native collagen fibrils. In addition, the measurements indicate a higher adhesion in the core of the collagen fibrils compared to the shell.

Authors:
 [1];  [2];  [1];  [2];  [1];  [2];  [1];  [2];  [3]
  1. Department of Geo- and Environmental Sciences, Ludwig-Maximilians-Universitaet, 80333 Munich (Germany)
  2. (Germany)
  3. GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg (Germany). E-mail: stefan.thalhammer@gsf.de
Publication Date:
OSTI Identifier:
20979812
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 354; Journal Issue: 1; Other Information: DOI: 10.1016/j.bbrc.2006.12.114; PII: S0006-291X(06)02759-8; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ATOMIC FORCE MICROSCOPY; COLLAGEN; ELASTICITY; IN VITRO; MORPHOLOGY; SPECTROSCOPY; YOUNG MODULUS

Citation Formats

Strasser, Stefan, GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg, Zink, Albert, GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg, Janko, Marek, GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg, Heckl, Wolfgang M., Deutsches Museum, Museumsinsel 1, 80538 Munich, and Thalhammer, Stefan. Structural investigations on native collagen type I fibrils using AFM. United States: N. p., 2007. Web. doi:10.1016/j.bbrc.2006.12.114.
Strasser, Stefan, GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg, Zink, Albert, GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg, Janko, Marek, GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg, Heckl, Wolfgang M., Deutsches Museum, Museumsinsel 1, 80538 Munich, & Thalhammer, Stefan. Structural investigations on native collagen type I fibrils using AFM. United States. doi:10.1016/j.bbrc.2006.12.114.
Strasser, Stefan, GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg, Zink, Albert, GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg, Janko, Marek, GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg, Heckl, Wolfgang M., Deutsches Museum, Museumsinsel 1, 80538 Munich, and Thalhammer, Stefan. Fri . "Structural investigations on native collagen type I fibrils using AFM". United States. doi:10.1016/j.bbrc.2006.12.114.
@article{osti_20979812,
title = {Structural investigations on native collagen type I fibrils using AFM},
author = {Strasser, Stefan and GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg and Zink, Albert and GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg and Janko, Marek and GSF-National Research Center for Environment and Health, Institute of Radiation Protection, AG Nanoanalytics, 85764 Neuherberg and Heckl, Wolfgang M. and Deutsches Museum, Museumsinsel 1, 80538 Munich and Thalhammer, Stefan},
abstractNote = {This study was carried out to determine the elastic properties of single collagen type I fibrils with the use of atomic force microscopy (AFM). Native collagen fibrils were formed by self-assembly in vitro characterized with the AFM. To confirm the inner assembly of the collagen fibrils, the AFM was used as a microdissection tool. Native collagen type I fibrils were dissected and the inner core uncovered. To determine the elastic properties of collagen fibrils the tip of the AFM was used as a nanoindentor by recording force-displacement curves. Measurements were done on the outer shell and in the core of the fibril. The structural investigations revealed the banding of the shell also in the core of native collagen fibrils. Nanoindentation experiments showed the same Young's modulus on the shell as well as in the core of the investigated native collagen fibrils. In addition, the measurements indicate a higher adhesion in the core of the collagen fibrils compared to the shell.},
doi = {10.1016/j.bbrc.2006.12.114},
journal = {Biochemical and Biophysical Research Communications},
number = 1,
volume = 354,
place = {United States},
year = {Fri Mar 02 00:00:00 EST 2007},
month = {Fri Mar 02 00:00:00 EST 2007}
}
  • Angiogenesis, or neovascularization, is tightly controlled by positive and negative regulators, many of which reside in the extracellular matrix. We have now identified eight novel 19- to 20-residue peptides derived from the {alpha}4, {alpha}5, and {alpha}6 fibrils of type IV collagen, which we have designated tetrastatins, pentastatins, and hexastatins, respectively. We have shown that these endogenous peptides suppress the proliferation and migration of HUVECs in vitro. By performing clustering analyses of the sequences using sequence similarity criteria and of the experimental results using a hierarchical algorithm, we report that the clusters identified by the experimental results coincide with the sequence-basedmore » clusters, indicating a tight relationship between peptide sequence and anti-angiogenic potency. These peptides may have potential as anti-angiogenic therapeutic agents.« less
  • No abstract prepared.
  • The Mov-13 mouse strain carries a retroviral insertion in the pro..cap alpha..1(I) collagen gene that prevents transcription of the gene. Cell lines derived from homozygous embryos do not express type I collagen although normal amounts of pro..cap alpha..2 mRNA are synthesized. The authors have introduced genomic clones of either the human or mouse pro..cap alpha..1(I) collagen gene into homozygous cell lines to assess whether the human or mouse pro..cap alpha..1(I) chains can associate with the endogenous mouse pro..cap alpha..2(I) chain to form stable type I collagen. The human gene under control of the simian virus 40 promoter was efficiently transcribedmore » in the transfected cells. Protein analyses revealed that stable heterotrimers consisting of two human ..cap alpha..1 chains and one mouse ..cap alpha..2 chain were formed and that type I collagen was secreted by the transfected cells at normal rates. However, the electrophoretic migration of both ..cap alpha..1(I) and ..cap alpha..2(I) chains in the human-mouse hybrid molecules were retarded, compared to the ..cap alpha..(I) chains in control mouse cells. Inhibition of the posttranslational hydroxylation of lysine and proline resulted in comigration of human and mouse ..cap alpha..1 and ..cap alpha..2 chains, suggesting that increased posttranslational modification caused the altered electrophoretic migration in the human-mouse hybrid molecules. Amino acid sequence differences between the mouse and human ..cap alpha.. chains may interfere with the normal rate of helix formation and increase the degree of posttranslational modifications similar to those observed in patients with lethal perinatal osteogenesis imperfecta. The Mov-13 mouse system should allow the authors to study the effect specific mutations introduced in transfected pro..cap alpha..1(I) genes have on the synthesis, assembly, and function of collagen I.« less
  • Collagen was labeled with )3,3,3-d/sub 3/) alanine and with (d/sub 10/) eucine via tissue culture. /sup 2/H nuclear magnetic resonance (NMR) spectra were obtained of collagen in solution and as fibrils using the quadrupolar echo techniqe. The /sup 2/H NMR data for (3,3,3-d/sub 3/)alanine-labeled collagen fibrils were analyzed in terms of a model for motion in which the molecule is considered to ump between two sites, separated azimuthally by an angle 2delta, in a time which is rapid compared with the residence time in both sites. The data suggest that the molecule undergoes reorientation over an angle, 2 delta, ofmore » approx. 30/sup 0/ in the fibrils, and that the average angle between the alanine C/sup ..cap alpha../-C/sup ..beta../ bond axis and the long axis of the helix is approx. 75/sup 0/. Reorientation is possibly segmental. The T/sub 2/ for (3,3,3-d/sub 3/)alanine-labeled collagen fibrils was estimated to be 105 ..mu..s. The /sup 2/H NMR data for the methyl groups of (d/sub 10/)leucine-labeled collagen were analyzed qualitatively. These data established that for collagen in solution and as fibrils, rotation occurs about the leucine side-chain bonds, in addition to threefold methyl rotation and reorientation of the peptide backbone. The T/sub 2/ for the methyl groups of leucine-labeled collagen is estimated to be approx. 130 ..mu..s. Taken together, these data provide strong evidence that both polypeptide backbone reorientation and amino acid side-chain motion occur in collagen molecules in the fibrils. Stabilizing interactions that determine fibril structure must therefore depend upon at least two sets of contacts in any given local region.« less