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Probing the biofunc...
Probing the biofunctionality of biotinylated hyaluronan and chondroitin sulfate by hyaluronidase degradation and aggrecan interaction
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- Altgärde, Noomi, 1983 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Nilebäck, Erik, 1984 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- De Battice, Laura, 1986 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Pashkuleva, I. (författare)
- Universidade do Minho,University of Minho
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- Reis, R. L. (författare)
- Universidade do Minho,University of Minho
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Becher, Jana (författare)
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Möller, Stephanie (författare)
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Schnabelrauch, Matthias (författare)
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- Svedhem, Sofia, 1970 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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(creator_code:org_t)
- Elsevier BV, 2013
- 2013
- Engelska.
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Ingår i: Acta Biomaterialia. - : Elsevier BV. - 1878-7568 .- 1742-7061. ; 9:9, s. 8158-8166
- Relaterad länk:
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http://dx.doi.org/10...
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https://research.cha...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Molecular interactions involving glycosaminoglycans (GAGs) are important for biological processes in the extracellular matrix (ECM) and at cell surfaces, and also in biotechnological applications. Enzymes in the ECM constantly modulate the molecular structure and the amount of GAGs in our tissues. Specifically, the changeable sulfation patterns of many GAGs are expected to be important in interactions with proteins. Biotinylation is a convenient method for immobilizing molecules to surfaces. When studying interactions at the molecular, cell and tissue level, the native properties of the immobilized molecule, i.e. its biofunctionality, need to be retained upon immobilization. Here, the GAGs hyaluronan (HA) and chondroitin sulfate (CS), and synthetically sulfated derivatives of the two, were immobilized using biotin-streptavidin binding. The degree of biotinylation and the placement of biotin groups (end-on/side-on) were varied. The introduction of biotin groups could have unwanted effects on the studied molecule, but this aspect that is not always straightforward to evaluate. Hyaluronidase, an enzyme that degrades HA and CS in the ECM, was investigated as a probe to evaluate the biofunctionality of the immobilized GAGs, using both quartz crystal microbalance and high-performance liquid chromatography. Our results showed that end-on biotinylated HA was efficiently degraded by hyaluronidase, whereas already a low degree of side-on biotinylation destroyed the degrading ability of the enzyme. Synthetically introduced sulfate groups also had this effect. Hence hyaluronidase degradation is a cheap and easy way to investigate how molecular function is influenced by the introduced functional groups. Binding experiments with the proteoglycan aggrecan emphasized the influence of protein size and surface orientation of the GAGs for in-depth studies of GAG behavior.
Ämnesord
- TEKNIK OCH TEKNOLOGIER -- Materialteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Materials Engineering (hsv//eng)
- NATURVETENSKAP -- Fysik -- Den kondenserade materiens fysik (hsv//swe)
- NATURAL SCIENCES -- Physical Sciences -- Condensed Matter Physics (hsv//eng)
Nyckelord
- Hyaluronidase
- Surface-sensitive techniques
- Glycosaminoglycan
- Biotin
- Aggrecan
Publikations- och innehållstyp
- art (ämneskategori)
- ref (ämneskategori)
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Till lärosätets databas
- Av författaren/redakt...
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Altgärde, Noomi, ...
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Nilebäck, Erik, ...
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De Battice, Laur ...
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Pashkuleva, I.
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Reis, R. L.
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Becher, Jana
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Möller, Stephani ...
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Schnabelrauch, M ...
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Svedhem, Sofia, ...
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- Om ämnet
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- TEKNIK OCH TEKNOLOGIER
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TEKNIK OCH TEKNO ...
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och Materialteknik
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- NATURVETENSKAP
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NATURVETENSKAP
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och Fysik
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och Den kondenserade ...
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Acta Biomaterial ...
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Chalmers tekniska högskola