ADAMTS4, His-Tag Active Protein | ADAMTS4 active protein

ADAMTS4, Recombinant, His-Tag (Aggrecanase I, A Disintegrin And Metalloproteinase with ThromboSpondin-3 motif)

Cat. Num. AAA638323

• Gene Names: ADAMTS4; ADMP-1; ADAMTS-2; ADAMTS-4
• Purity: Purified; 90%. Appears as a major band of 42kD in SDS-PAGE. Due to autoproteolytic activity minor bands of ADAMTS4 may be visiable in the enzymatic preparation.
• Synonyms: ADAMTS4; His-Tag; Recombinant; His-Tag (Aggrecanase I; A Disintegrin And Metalloproteinase with ThromboSpondin-3 motif); ADAMTS4 active protein

• Purity/Purification: Purified; 90%. Appears as a major band of 42kD in SDS-PAGE. Due to autoproteolytic activity minor bands of ADAMTS4 may be visiable in the enzymatic preparation.
• Form/Format: Supplied as a liquid in 50mM Tris-HCl, pH 7.5, 150mM sodium chloride, 5mM calcium chloride, 0.05% Brij-35.
• Sequence: The protein consists of amino acids F213-A579 of full-length ADAMTS4 and a C-terminal His6tag: FASLSRFVETLVVADDKMAAFHG AGLKRYLLTVMAAAAKAFKHPSIRNPVSLV VTRLVILGSGEEGPQVGPSAAQTLRSFCA WQRGLNTPEDSDPDHFDTAILFTRQDLC GVSTCDTLGMADVGTVCDPARSCAIVEDD GLQSAFTAAHELGHVFNMLHDNSKPCISL NGPLSTSRHVMAPVMAHVDPEEPWSPCS ARFITDFLDNGYGHCLLDKPEAPLHLPVTF GKDYDADRQCQLTFGPDSRHCPQLPPP CAALWCSGHLNGHAMCQTKHSPWADG TPCGPAQACMGGRCLHMDQLQDFNIPQ AGGWGPWGPWGDCSRTCGGGVQFSSR DCTRPVPRNGGKYCEGRRTRFRSCNTEDC PTGSA-(H)6.

• Application Notes: Recombinant ADAMTS4 is used to study the degradation of extracellular matrix proteoglycans, to screen for inhibitors of proteoglycan hydrolysis and to characterize inhibitor actions. The enzyme can also serve as standard in enzymatic and immunochemical assays.
• Activity: Aggrecanase activity of ADAMTS4 is determined with recombinant His-tagged aggrecan interglobular domain. ADAMTS4 hydrolyzes the "aggrecanase" site within this domain (peptide bond E373-A374 in human aggrecan). The recombinant substrate is incubated at a concentration of 1uM with 5nM ADAMTS4 in 50mM Tris-HCI, pH 7.5, 150mM sodium chloride, 5mM calcium chloride, 1uM leupeptin, 1uM pepstatin, 1mM PMSF, 0.02% Brij 35 for 1hr at 37 degree C. Cleavage at the "aggrecanase" site is estimated from the appearance of the C-terminus ARGSVIL. Using polyclonal neoepitope-antibodies to the ARGS-terminus the fragment is fixed to a microplate and quantified with anti-His-tag antibody. Under the specified conditions recombinant ADAMTS4 hydrolyzes 0.1nmoles substrate/ml • h. The specific activity is accordingly 1.8 nmoles hydrolyzed substrate/min • mg enzyme. Activities of different enzyme lots may vary by up to 50%.
• Preparation and Storage: Aliquot to avoid repeated freezing and thawing and freeze at -70 degree C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquots are stable for at least 12 months.

SDS-Page

(SDS-PAGE of 0.5ug of ADAMTS4)

Related Product Information for ADAMTS4 active protein

ADAMTS (a disintegrin and metalloproteinase with thrombospondin motif) is a novel family of extrcellular proteinases (1). Presently nine family members have been identified in mammals (1). ADAMTS4 was first purified from I1-1 (stimulated bovine nasal cartilage conditioned media) and human ADAMTS4 (cDNA was cloned from a human heart cDNA library) (2). Mature ADAMTS4 consists of a prodomain which confers latency to the proenzyme, a catalytic domain, a disintegrin domain and a C-terminal sequence with a thrombospondin type-1 motif. The prodomain is most likely cleaved off by a furin-type enzyme before active ADAMTS4 is released from cells. Active ADAMTS4 consists of 625 amino acids (F213-K837) with a calculated Mr of 67 943 (2).ADAMTS4 hydrolyzes aggrecan, the major proteoglycan of articular cartilage (2). As aggrecan is also digested by 2 other members of the ADAMTS family, ADAMTS1(3) and ADAMTS5 (4), aggrecan degradation products found in normal and rheumatoid and osteoarthritic joint cartilage (5) may arise from action of either of the 3 proteinases. Isolated ADAMTS4 hydrolyzes aggrecan at 5 different sites (6). Four cleavage sites are located in the chondroitin sulfate-rich region between globular domains G2 and G3, while one site is placed in the rod-shaped polypeptide between globular domains G1 and G2. The thrombospondin motif in ADAMTS4 appears to be critical for aggrecan substrate recognition and cleavage (7). ADAMTS4 hydrolyzes also other lecticans as brevican (8) and versican (9).

Product Categories/Family for ADAMTS4 active protein

Molecular Biology; MB-Matrix Metalloproteinases

NCBI and Uniprot Product Information

• NCBI GI #: 157427675
• NCBI GeneID: 9507
• NCBI Accession #: NP_005090.3
• NCBI GenBank Nucleotide #: NM_005099.4
• UniProt Accession #: O75173; Q5VTW2; Q6P4Q8; Q6UWA8; Q9UN83
• Molecular Weight: 90,197 Da
• NCBI Official Full Name: A disintegrin and metalloproteinase with thrombospondin motifs 4 preproprotein
• NCBI Official Synonym Full Names: ADAM metallopeptidase with thrombospondin type 1 motif, 4
• NCBI Official Symbol: ADAMTS4
• NCBI Official Synonym Symbols: ADMP-1; ADAMTS-2; ADAMTS-4
• NCBI Protein Information: A disintegrin and metalloproteinase with thrombospondin motifs 4; ADAM-TS4; ADAM-TS 4; aggrecanase-1; a disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif, 4
• UniProt Protein Name: A disintegrin and metalloproteinase with thrombospondin motifs 4
• Protein Family: A disintegrin and metalloproteinase with thrombospondin motifs
• UniProt Gene Name: ADAMTS4
• UniProt Synonym Gene Names: KIAA0688; ADAM-TS 4; ADAM-TS4; ADAMTS-4
• UniProt Entry Name: ATS4_HUMAN

NCBI Description

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 lacks a C-terminal TS motif. It is responsible for the degradation of aggrecan, a major proteoglycan of cartilage, and brevican, a brain-specific extracellular matrix protein. The cleavage of aggrecan and brevican suggests key roles of this enzyme in arthritic disease and in the central nervous system, potentially, in the progression of glioma. [provided by RefSeq, Jul 2008]

Uniprot Description

ADAMTS4: 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. Could also be a critical factor in the exacerbation of neurodegeneration in Alzheimer disease. Cleaves aggrecan at the '392-Glu-|-Ala-393' site.

Protein type: Motility/polarity/chemotaxis; Secreted, signal peptide; EC 3.4.24.82; Secreted; Protease

Chromosomal Location of Human Ortholog: 1q21-q23

Cellular Component: extracellular matrix; extracellular space; proteinaceous extracellular matrix; extracellular region

Molecular Function: peptidase activity; protein binding; protease binding; metallopeptidase activity; zinc ion binding; metalloendopeptidase activity

Biological Process: extracellular matrix disassembly; extracellular matrix organization and biogenesis; defense response to bacterium; proteolysis; skeletal development