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描述 产品目录编号 CC1034 品牌系列 Chemicon®
商名 - Chemicon
描述 ADAMTS-5, recombinant human truncated 概述 ADAMTS's (a disintegrin and metalloproteinase with thrombospondin motif) are multidomain extracellular proteinases, containing at least one thrombospondin type-1 repeat (Tang, 2001). The enzymes are involved in processing of extracellular matrix proteins and proteins in the circulation. ADAMTS-5 was first purified from bovine nasal cartilage conditioned media and human ADAMTS-5 cDNA was cloned from a human liver cDNA library (Abbaszade, I. et al, 1999). ADAMTS-5 consists of a prodomain, a catalytic domain, a disintegrin domain, a thrombospondin type-1 motif, a Cys-rich region followed by a spacer sequence and thrombospondin submotifs. The prodomain is most likely cleaved-off by a furin-type enzyme before ADAMTS-5 is released from cells. ADAMTS-5 is expressed constitutively in synovium (Vankemmelbeke, MN. et al, 2001) and it degrades aggrecan, the major proteoglycan of articular cartilage. Like ADAMTS4, another member of the ADAMTS-family, ADAMTS-5 cleaves aggrecan at 5 sites (Tortorella, MD. et al, 2002). Four cleavage sites are located in the chondroitin sulfate-rich region between aggrecan globular domains G2 and G3, while one site is contained in the rod-shaped polypeptide between globular domains G1 and G2. In addition, ADAMTS-5 cleaves one more site in the chondroitin sulfate-rich region that is not cleaved by ADAMTS4 (Tortorella, MD. et al, 2002). ADAMTS-5 is inhibited by TIMP 3 (Kashigawa, M. et al, 2001) and a2-macroglobulin (Tortorella, MD. et al, 2004).
Molecular form: Recombinant human ADAMTS-5 D625-930 is produced with the baculo-virus expression system and purified from insect cell culture supernatants. The protein contains the catalytic domain, the disintegrin domain and the thrombospondin type-1 motif of full-length ADAMTS-5 followed by a C-terminal His6-tag. The calculated Mr of the amino acid sequence is 40.8 kDa. In SDS-PAGE the protein exhibits a Mr of about 50 kDa.
ACTIVITY:
Aggrecanase activity of the ADAMTS-5 preparation is determined with recombinant aggrecan interglobular domain (Aggrecan-IGD from Chemicon). ADAMTS-5 hydrolyzes the "aggrecanase" site within this domain (peptide bond E373 -A374 in human aggrecan). The recombinant substrate is incubated at a concentration of 0.1 μM with ADAMTS-5 in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 1 μM leupeptin, 1 μM pepstatin, 1 mM Pefabloc, 0.05 % Brij 35 for 15 min at 37 °C. Substrate cleavage at the "aggrecanase"-site is estimated from the appearance of the hydrolysis fragment with the novel N-terminus ARGSVIL. The fragment is quantified with two monoclonal antibodies. Under the specified conditions the hydrolysis rate is > 0.5 nmoles hydrolyzed substrate/ min . ml ADAMTS-5 preparation or > 5 nmoles hydrolyzed substrate/ min . mg.
Inhibitors: ADAMTS-5 is inhibited by tissue inhibitor of matrix metalloproteinase 3 (TIMP3) and by alpha2-macroglobulin. Enzyme activity is also suppressed by chelators of divalent cations such as EDTA and by synthetic metalloproteinase inhibitors.Alternate Names - Aggrecanase 2
产品信息 演示 Solubilized in 50mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 50mM imidazol 0.05 % Brij-35. 应用 主要应用 - Affects Function
应用说明 Functional Studies
Degradation of extracellular matrix proteoglycans
Screening and characterisation of inhibitors
Standard in enzymatic and immunological assays
Optimal working dilutions must be determined by the end user.生物信息 浓缩 0.1 mg/mL 纯度 As judged from SDS-PAGE, truncated ADAMTS-5 represents > 50% of total protein of the preparation. The total protein concentration is 0.1 mg/ml. 品种 Human Entrez基因编号 - NM_007038.2
Entrez基因汇总 This gene encodes a member of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) protein family. Members of the family share several distinct protein modules, including a propeptide region, a metalloproteinase domain, a disintegrin-like domain, and a thrombospondin type 1 (TS) motif. Individual members of this family differ in the number of C-terminal TS motifs, and some have unique C-terminal domains. The enzyme encoded by this gene contains two C-terminal TS motifs and functions as aggrecanase to cleave aggrecan, a major proteoglycan of cartilage. 基因符号 - EC 3.4.24.-
- ADAMTS5
- ADAM-TS5
- ADAMTS11
- Aggrecanase-2
- ADMP2
- ADMP-2
- FLJ36738
- aggrecanase-2
UniProt编号 - Q9UNA0
UniProt汇总 FUNCTION: SwissProt: Q9UNA0 # Cleaves aggrecan, a cartilage proteoglycan, and may be involved in its turnover. May play an important role in the destruction of aggrecan in arthritic diseases. May play a role in proteolytic processing mostly during the peri-implantation period.
COFACTOR: Binds 1 zinc ion per subunit (By similarity).
SIZE: 930 amino acids; 101716 Da
SUBCELLULAR LOCATION: Secreted, extracellular space, extracellular matrix (By similarity).
TISSUE SPECIFICITY: Expressed at low level, primarily in placenta but also in other tissues, such as heart and brain, and also cervix, uterus, bladder, esophagus, rib cartilage, chondroblastoma, fibrous tissue and joint capsule from an arthritic patient.
DOMAIN: SwissProt: Q9UNA0 The spacer domain and the TSP type-1 domains are important for a tight interaction with the extracellular matrix. & The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme.
PTM: The precursor is cleaved by a furin endopeptidase (By similarity).
SIMILARITY: Contains 1 disintegrin domain. & Contains 1 peptidase M12B domain. & Contains 2 TSP type-1 domains.产品使用声明 使用声明 - Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
储存和货运信息 存储条件 Maintain at -70°C for 12 months from date of receipt. The enzyme can be kept at -20°C for several weeks and ice for several days. Avoid repeated freeze/thaw cycles. 包装信息 数量 5 µg
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