Recombinant Hu ADAMTS-5

Description
Catalogue Number	CC1034
Brand Family	Chemicon®
Trade Name	Chemicon
Description	ADAMTS-5, recombinant human truncated
Overview	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

Description

Catalogue Number

CC1034

Brand Family

Chemicon®

Trade Name

Chemicon

Description

ADAMTS-5, recombinant human truncated

Overview

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

Product Information
Presentation	Solubilized in 50mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 50mM imidazol 0.05 % Brij-35.

Product Information

Presentation

Solubilized in 50mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl2, 50mM imidazol 0.05 % Brij-35.

Applications
Key Applications	Affects Function
Application Notes	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.

Applications

Key Applications

Affects Function

Application Notes

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.

Biological Information
Concentration	0.1 mg/mL
Purity	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.
Entrez Gene Number	NM_007038.2
Entrez Gene Summary	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.
Gene Symbol	ADAMTS5 ADAM-TS5 ADAMTS11 Aggrecanase-2 ADMP2 ADMP-2 FLJ36738 aggrecanase-2 EC 3.4.24.-
UniProt Number	Q9UNA0
UniProt Summary	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.

Biological Information

Concentration

0.1 mg/mL

Purity

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.

Entrez Gene Number

NM_007038.2

Entrez Gene Summary

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.

Gene Symbol

ADAMTS5
ADAM-TS5
ADAMTS11
Aggrecanase-2
ADMP2
ADMP-2
FLJ36738
aggrecanase-2
EC 3.4.24.-

UniProt Number

Q9UNA0

UniProt Summary

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.

Product Usage Statements
Usage Statement	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.

Product Usage Statements

Usage Statement

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.

Storage and Shipping Information
Storage Conditions	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.

Storage and Shipping Information

Storage Conditions

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.

Packaging Information
Material Size	5 µg

Packaging Information

Material Size

5 µg

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Recombinant Hu ADAMTS-5

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