title: Detection of ADAMTS-4 activity using a fluorogenic peptide-conjugated Au nanoparticle probe in human knee synovial fluid.
abstract: A disintegrin and metalloproteinase with thrombospondin motif-4 (ADAMTS-4) plays a pivotal role in degrading aggrecan, which is an early event in cartilage degrading joint diseases such as osteoarthritis (OA). Detection of ADAMTS-4 activity could provide useful clinical information for early diagnosis of such diseases and disease-modifying therapy. Therefore, we developed a ADAMTS-4 detective fluorescent turn-on AuNP probe (ADAMTS-4-D-Au probe) by conjugating gold nanoparticles with a FITC-modified ADAMTS-4-specific peptide (DVQEFRGVTAVIR). When the ADAMTS-4-D-Au probe was incubated with ADAMTS-4, the fluorescence recovered and fluorescence intensity markedly increased in proportion to concentrations of ADAMTS-4 and the probe. A nearly 3-fold increase in fluorescent intensity in response to only 3.9 pM of ADAMTS-4 was detected, whereas almost no fluorescence recovery was observed when the probe was incubated with matrix metalloproteinase (MMP)-1, -3, and -13. These results indicate a relative high sensitivity and specificity of the probe. Moreover, ADAMTS-4-D-Au probe was used to detect ADAMTS-4 activity in synovial fluid from 11 knee surgery patients. A substantial increase in fluorescent intensity was observed in the acute joint injury group as compared to the chronic joint injury and end-stage OA groups, indicating that this simple and low-cost sensing system might serve as a new detection method for ADAMTS-4 activity in biological samples and in screens for inhibitors for ADAMTS-4-related joint diseases. Additionally, this probe could be a potential biomarker for early diagnosis of cartilage-degrading joint diseases.
authors.affiliation: Institute of Sports Medicine, Peking University Third Hospital, Haidian District, Beijing, P R China.
authors.forename: Shi
authors.initials: S
authors.lastname: Peng
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zheng
authors.forename: Xin
authors.initials: X
authors.lastname: Zhang
authors.forename: Linghui
authors.initials: L
authors.lastname: Dai
authors.forename: Jingxian
authors.initials: J
authors.lastname: Zhu
authors.forename: Yanbin
authors.initials: Y
authors.lastname: Pi
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Hu
authors.forename: Wenqing
authors.initials: W
authors.lastname: Cheng
authors.forename: Chunyan
authors.initials: C
authors.lastname: Zhou
authors.forename: Yinlin
authors.initials: Y
authors.lastname: Sha
authors.forename: Yingfang
authors.initials: Y
authors.lastname: Ao
mesh_terms: D051722:ADAM Proteins / Q000032:analysis* / Q000378:metabolism; D000071121:ADAMTS4 Protein; D057075:Enzyme Assays / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D006801:Humans; D007719:Knee Joint / Q000737:chemistry / Q000201:enzymology*; D053758:Nanoparticles / Q000737:chemistry*; D010003:Osteoarthritis / Q000175:diagnosis / Q000201:enzymology*; D010455:Peptides / Q000737:chemistry; D011348:Procollagen N-Endopeptidase / Q000032:analysis* / Q000378:metabolism; D013582:Synovial Fluid / Q000737:chemistry / Q000201:enzymology*
chemical_list: D005456:Fluorescent Dyes; D010455:Peptides; D006046:Gold; D051722:ADAM Proteins; D011348:Procollagen N-Endopeptidase; D000071121:ADAMTS4 Protein; C000603918:ADAMTS4 protein, human