Neurocan-GFP fusion protein: a new approach to detect hyaluronan on tissue sections and living cells.
Hyaluronan is an unsulfated glycosaminoglycan (GAG) that is ubiquitously expressed in the extracellular matrix (ECM) of all vertebrates, where hyaluronan rich matrices constitute a particular permissive environment for the development of complex biological structures and also for tumor progression. Because of its conserved structure and ubiquitous expression, antibodies for its histochemical detection cannot be produced. We have engineered a fusion protein, neurocan-GFP, and expressed it as a secreted molecule in mammalian cells. Neurocan-GFP fusion protein specifically binds to hyaluronan and directly visualizes hyaluronan on tissue sections, revealing a very detailed picture of hyaluronan distribution. The fluorescent fusion protein can be used in combination with antibodies and nuclear markers for double or triple staining. In addition, it is suitable to visualize hyaluronan on living cells by time-lapse video microscopy. The successful production and application of the neurocan-GFP fusion protein opens up new perspectives for using GFP fusion proteins as detection tools in histological and cytological studies complementing conventional antibody and biotin/avidin techniques.
Zhang
Hui
H
Department of Experimental Pathology, Lund University, Lund, Sweden.
Baader
Stephan L
SL
Sixt
Michael
M
Kappler
Joachim
J
Rauch
Uwe
U
D000818:Animals; D000831:Animals, Newborn; D002460:Cell Line; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics*; D004285:Dogs; D005123:Eye / Q000033:anatomy & histology / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism*; D037181:Lectins, C-Type; D008164:Luminescent Proteins / Q000235:genetics*; D051379:Mice; D008856:Microscopy, Fluorescence; D009419:Nerve Tissue Proteins / Q000235:genetics*; D058580:Neurocan; D011485:Protein Binding; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014741:Video Recording
D011508:Chondroitin Sulfate Proteoglycans; D037181:Lectins, C-Type; D008164:Luminescent Proteins; D009419:Nerve Tissue Proteins; D058580:Neurocan; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C076817:NCAN protein, human; D006820:Hyaluronic Acid