Please use this identifier to cite or link to this item: http://hdl.handle.net/1822/24875

TitleProbing the biofunctionality of biotinylated hyaluronan and chondroitin sulfate by hyaluronidase degradation and aggrecan interaction
Author(s)Altgärde, Noomi
Nilebäck, Erik
Battice, Laura de
Pashkuleva, I.
Reis, R. L.
Becher, Jana
Möller, Stephanie
Schnabelrauch, Matthias
Svedhem, Sofia
KeywordsGlycosaminoglycan
Biotin
Surface-sensitive techniques
Hyaluronidase
Aggrecan
Issue dateJun-2013
PublisherElsevier
JournalActa Biomaterialia
Abstract(s)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.
TypeArticle
DescriptionAvailable online 5 June 2013
URIhttp://hdl.handle.net/1822/24875
DOI10.1016/j.actbio.2013.05.031
ISSN1742-7061
Publisher versionhttp://dx.doi.org/10.1016/j.actbio.2013.05.031
Peer-Reviewedyes
AccessRestricted access (UMinho)
Appears in Collections:3B’s - Artigos em revistas/Papers in scientific journals

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