Supercharged fluorescent protein as a versatile probe for the detection of glycosaminoglycans in vitro and in vivo.
Glycosaminoglycans (GAGs) are linear acidic heteropolysaccharides that are ubiquitously expressed in animal tissues and participate in various life processes. To date, the detection and visualization of GAGs in complex biological samples and living organisms remain a challenge because of the lack of powerful biocompatible probes. In this study, a superpositively charged green fluorescent protein (ScGFP) was shown great potential in GAG detection for the first time. First, on the basis of the phenomenon of GAGs dose-dependently inhibiting the fluorescence quenching of ScGFP by graphene oxide, a simple and highly sensitive signal-on homogeneous platform was established for detecting and quantifying GAGs, even in complex samples such as heparin in citrated plasma and oversulfated chondroitin sulfate in heparin. Furthermore, ScGFP with excellent stability and biocompatibility could be easily used as a highly sensitive and selective probe to visualize different types of GAGs in vitro and in vivo through combination with specific GAG-degrading enzymes. This study introduces a versatile probe for GAG detection, which is easy to prepare and which shows a high practical value in basic research and medical applications.
National Glycoengineering Research Center and State Key Laboratory of Microbial Technology, Shandong University , Jinan, Shandong 250100, P. R. China.
Wenshuang
W
Wang
National Glycoengineering Research Center and State Key Laboratory of Microbial Technology, Shandong University , Jinan, Shandong 250100, P. R. China.
Naihan
N
Han
Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology , Qingdao, Shandong 266042, P. R. China.
Ruijuan
R
Li
National Glycoengineering Research Center and State Key Laboratory of Microbial Technology, Shandong University , Jinan, Shandong 250100, P. R. China.
Wenjun
W
Han
Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology , Qingdao, Shandong 266042, P. R. China.
Xiaoru
X
Zhang
National Glycoengineering Research Center and State Key Laboratory of Microbial Technology, Shandong University , Jinan, Shandong 250100, P. R. China.
Fuchuan
F
Li
D000818:Animals; D002236:Carbohydrate Conformation; D002417:Cattle; D002460:Cell Line; D002470:Cell Survival; D019343:Citric Acid / Q000737:chemistry; D006025:Glycosaminoglycans / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D015335:Molecular Probes / Q000737:chemistry* / Q000378:metabolism
D006025:Glycosaminoglycans; D015335:Molecular Probes; D049452:Green Fluorescent Proteins; D019343:Citric Acid