ADAM Metallopeptidase with Thrombospondin Type 1 Motif, 4 (ADAMTS4) Protein

Name: ADAM Metallopeptidase with Thrombospondin Type 1 Motif, 4 (ADAMTS4)

Synonyme: ADMP-1, ADAMTS-2, ADAMTS-4, KIAA0688, KIAA0366, ADAM-TS4, mKIAA0688, 3830423K05, ADAMTS4, DKFZp469J0817, AAC2, PNAT, NPI, PCINP, PCPNI, hPCPNI, ADAM-TS2, ADAMTS-3, TTP, VWFCP, C9orf8, vWF-CP, FLJ42993, MGC118899, MGC118900, DKFZp434C2322, ADMP-2, ADAMTS11, FLJ36738, ASMP-2, ADAMTS1, ADAM-TS5, AI481094, 9530092O11Rik, PWTSR, THSD2, CRISTIN1, FLJ14440, AW743315, 1810011L16Rik, 8430403M15Rik, E030027K14Rik, FLJ16042, AU015154, 4831407I23Rik, D230034E10Rik, Gm710, ADAMTS5, ADAMTS13, ADAMTS-11

Herkunft (Gen): Human

Applikation: Blockierende Antikörper (Inhibition), ELISA

Produktnummer: ABIN368661

Produktbeschreibung

Produktmerkmale: Recombinant Human ADAMTS4 (Aggrecanase 1) Truncated, His-tagged

Beschreibung: Molecular form: Recombinant human ADAMTS4 delta580-837 is produced with the baculovirus expression system and purified from insect cell culture supernatants. The protein consists of amino acids F 213 .A 579 of full-length ADAMTS4 and a C-terminal His 6 -tag: F A S L S R F V E T L V V A D D K M A A F H G A G L K R Y L L T V M A A A A K A F K H P S I R N P V S L V V T R L V I L G S G E E G P Q V G P S A A Q T L R S F C A W Q R G L N T P E D S D P D H F D T A I L F T R Q D L C G V S T C D T L G M A D V G T V C D P A R S C A I V E D D G L Q S A F T A A H E L G H V F N M L H D N S K P C I S L N G P L S T S R H V M A P V M A H V D P E E P W S P C S A R F I T D F L D N G Y G H C L L D K P E A P L H L P V T F P G K D Y D A D R Q C Q L T F G P D S R H C P Q L P P P C A A L W C S G H L N G H A M C Q T K H S P W A D G T P C G P A Q A C M G G R C L H M D Q L Q D F N I P Q A G G W G P W G P W G D C S R T C G G G V Q F S S R D C T R P V P R N G G K Y C E G R R T R F R S C N T E D C P T G S A (H) 6 The calculated M r is 40 366 Da. ADAMTS4 is solublized in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl 2 , 0.05 % Brij-35. Specific activity: Aggrecanase activity of ADAMTS4 is determined with recombinant aggrecan interglobular domain (Aggrecan-IGD from Invitek). ADAMTS4 hydrolyzes the "aggrecanase" site within this domain (peptide bond E 373 -A 374 in human aggrecan). The recombinant substrate is incubated at a concentration of 0.1 uM with 1 nM ADAMTS4 in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 5 mM CaCl 2 , 1 uM leupeptin, 1 uM pepstatin, 1 mM Pefabloc, 0.05 % Brij 35 for 15 min at 37 o C. 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, one directed against the neoepitope ARGSVIL, the other against the sequence C-terminal from the neoepitope (InviLISA Aggrecanase Activity Assay). Under the specified conditions the hydrolysis rate is > 0.15 nM hydrolysed substrate/ min x nM truncated ADAMTS4. When related to mg enzyme the value is > 3.7 nmoles hydrolyzed substrate/ min x mg ADAMTS4. 5040 30 20 kDa 5040 30 20 5040 30 20 5040 30 20 5040 30 20 kDa SDS-PAGE of 0.5 u g ADAMTS4 . Inhibitors: ADAMTS4 is inhibited by tissue inhibitor of matrix metalloproteinases 3 (TIMP 3) and to a smaller extent by other tissue inhibitors. Enzyme activity is also suppressed by chelators of divalent cations as EDTA and by synthetic metalloproteinase inhibitors. Strucutre and function: ADAMTS4 ADAMTS (a disintegrin and metalloproteinase with thrombospondin motif) is a novel family of extracellular proteinases [1]. Presently nineteen family members have been identified in the human genome. ADAMTS4 was first purified from Il-1-stimulated bovine nasal cartilage conditioned media and human ADAMTS4- cDNA was cloned from a human heart cDNA library [2]. ADAMTS4 consists of a prodomain, a catalytic domain, a disintegrin domain, a thrombospondin type-1 motif and a Cys-rich region followed by a C- terminal sequence. The prodomain is most likely cleaved off by a furin-type enzyme before ADAMTS4 is released from cells. 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 any 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]. ADAMTS4 is inhibited by TIMP 3 [10] and by the N-terminal domain of TIMP 3 [11] with K i -values in the nanomolar range. Inhibition by TIMP 1, 2 and 4 is much weaker [10].

Synonyme: ADMP-1, ADAMTS-2, ADAMTS-4, KIAA0688, KIAA0366, ADAM-TS4, mKIAA0688, 3830423K05, ADAMTS4, DKFZp469J0817, AAC2, PNAT, NPI, PCINP, PCPNI, hPCPNI, ADAM-TS2, ADAMTS-3, TTP, VWFCP, C9orf8, vWF-CP, FLJ42993, MGC118899, MGC118900, DKFZp434C2322, ADMP-2, ADAMTS11, FLJ36738, ASMP-2, ADAMTS1, ADAM-TS5, AI481094, 9530092O11Rik, PWTSR, THSD2, CRISTIN1, FLJ14440, AW743315, 1810011L16Rik, 8430403M15Rik, E030027K14Rik, FLJ16042, AU015154, 4831407I23Rik, D230034E10Rik, Gm710, ADAMTS5, ADAMTS13, ADAMTS-11

Anwendungen

Applikationshinweise: Recombinant ADAMTS4 is used to study the degradation of extracellular matrix proteoglycans, to screen for inhibitors of proteoglycan hydrolysis and to characterise inhibitor actions. The enzyme can also serve as standard in enzymatic and immunochemical assays.

Reinheit: Recombinant ADAMTS4 appears as a major protein of about 42 kDa in SDS-PAGE ( > 90 % of total protein). Due to autoproteolytic activity minor bands of ADAMTS4 may be visible in the enzyme preparation.

Lagerung: Recombinant ADAMTS4 is stable until the expiry date given on the label when stored at -70°C. The enzyme can be kept at -20°C for several weeks. Repeated freezing and thawing should be avoided.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Bilder

ADAM Metallopeptidase with Thrombospondin Type 1 Motif, 4 (ADAMTS4) protein