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CC1890 Aggrecan Interglobular Domain, Recombinant, His-tagged

CC1890
50 µg  
Purchase on Sigma-Aldrich

Áttekintés

Replacement Information
Description
Catalogue NumberCC1890
Brand Family Chemicon®
Trade Name
  • Chemicon
DescriptionAggrecan Interglobular Domain, Recombinant, His-tagged
OverviewMolecular form: The polypeptide connecting human aggrecan globular domains 1 and 2 (T331 - G458) is expressed in E. coli with a C-terminal twin His-tag {6His-L-E-6His}. The recombinant protein contains cleavage sites for aggrecanases (E373 - A374) and matrix metalloproteinases (N341 - F342). It comprises the following amino acids: T A E D F V D I P E N<> F F G V G G E E D I T V Q T V T W P D M E L P L P R N I T E G E <>A R G S V I L T V K P I F E V S P S P L E P E E P F T F A P E I G A T A F A E V E N E T G E A T R P W G F P T P G L G P A T A F T S E D L V V Q V T A V P G Q P H L P G G (His-tag)

Main cleavage sites are indicated by <> inclusions.. The calculated Mr of the His-tagged protein is 15,493 Da.

Purity: The recombinant aggrecan interglobular domain appears as a major band at about 21 kDa in SDS-PAGE. It represents more than 90% of total protein in the preparation.

Applications: Aggrecan interglobular domain is used as substrate for aggrecanases and matrix metalloproteinases. For proteinase activity measurements, the protein is incubated with proteinase for various time intervals. Thereafter, aliquots of the incubation mixture are analyzed by SDS-PAGE or by ELISA.Upon cleavage with aggrecanases the apparent Mr of aggrecan interlobular domain in SDS-PAGE is reduced from 21 kDa to about 13 kDa. Quantitative measurement of aggrecanase cleavage requires a neoepitop antibody with specificity for the N-terminus A R G S V I L T . . . appearing upon hydrolysis. The fragment with the newly formed N-terminus is fixed by the neoepitop antibody to a microplate and quantified with an anti-His-tag antibody. In analogy, cleavage by matrix metalloproteinases can be measured with antibodies to neoepitopes appearing upon action of these enzymes.
Alternate Names
  • Aggrecan-IGD1
Background InformationAggrecan is a large aggregating proteoglycan of articular cartilage. It is also found in aorta, discs and tendons [Knudsen & Knudsen, 2001; Watanabe et al., 1998]. The aggrecan core protein consists of 2317 amino acids [Watanabe et al., 1998]. Up to 130 glucosaminoglycan chains are attached to the core protein and the total molecular mass can reach 2.2 - 3.0 x 106 Daltons [Hardingham & Fosang, 1992]. Within the aggrecan molecule 3 global domains G1, G2 and G3 can be distinguished. Domains G1 and G2 are connected by a rod-shaped polypeptide called interglobular domain (IGD), while the sequence between domains G2 and G3 contains attachment regions for keratan sulfate and chondroitin sulfate chains. Aggrecan interacts via the G1domain with hyaluronan and link protein to form large aggregates. Such aggregates can contain up to 50-100 aggrecan monomers noncovalently bound to a single hyaluronan chain through 2 link proteins [Knudsen & Knudsen, 2001; Watanabe et al., 1998; Hardingham & Fosang, 1992]. The aggregates form a hydrated gel-like structure, which endowes cartilage with resistibility to compression and deformation. Degradation of aggrecan appears to initiate at the C-terminus. The population of aggrecan molecules without the G3 domain increases with aging [Dudhia et al., 1996]. Isolated aggrecanases cleave aggrecan at 4 sites within the chrondroitin sulfate-rich region (sites E1667 -G1668, E1480-G1481, E1771 - A1772, E1871 - L1872) and 1 site within the interglobular domain (E373 - A374) [Tortorella et al., 2000]. Cleavage at the latter site had been documented by analysis of cartilage proteoglycan breakdown products in rheumatoid and osteoarthritis [Lohmander et al., 1993]. To measure aggrecanase activity, an artificial recombinant protein composed of aggrecan interglobular domain with flanking FLAG-sequence and human immunoglobulin G1 constant region was first used by Hughes et al. [Hughes et al., 1997].
References
Product Information
PresentationThe calculated Mr of the His-tagged protein is 15,493 Da. The protein is solubilized in 50 mM Tris-HCI, pH 7.5, 150 mM NaCl, 5 mM CaCl2.
Quality LevelMQ100
Applications
Key Applications
  • Affects Function
Biological Information
SourceHuman
Entrez Gene Number
Entrez Gene SummaryThis gene is a member of the aggrecan/versican proteoglycan family. The encoded protein is an integral part of the extracellular matrix in cartilagenous tissue and it withstands compression in cartilage. Mutations in this gene may be involved in skeletal dysplasia and spinal degeneration. Multiple alternatively spliced transcript variants that encode different protein isoforms have been observed in this gene.
Gene Symbol
  • ACAN
  • AGC1
  • MSK16
  • CSPG1
  • SEDK
  • CSPCP
  • AGCAN
  • CSPGCP
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P16112 # This proteoglycan is a major component of extracellular matrix of cartilagenous tissues. A major function of this protein is to resist compression in cartilage. It binds avidly to hyaluronic acid via an N-terminal globular region.
SIZE: 2415 amino acids; 250193 Da
SUBUNIT: Interacts with FBLN1 (By similarity).
SUBCELLULAR LOCATION: Secreted, extracellular space, extracellular matrix (By similarity).
TISSUE SPECIFICITY: Restricted to cartilages.
DEVELOPMENTAL STAGE: Expression was detected in chondrocytes throughout the developing skeleton.
DOMAIN: SwissProt: P16112 Two globular domains, G1 and G2, comprise the N-terminus of the proteoglycan, while another globular region, G3, makes up the C-terminus. G1 contains Link domains and thus consists of three disulfide-bonded loop structures designated as the A, B, B' motifs. G2 is similar to G1. The keratan sulfate (KS) and the chondroitin sulfate (CS) attachment domains lie between G2 and G3.
PTM: Contains mostly chondroitin sulfate, but also keratan sulfate chains, N-linked and O-linked oligosaccharides. The release of aggrecan fragments from articular cartilage into the synovial fluid at all stages of human osteoarthritis is the result of cleavage by aggrecanase.
DISEASE: SwissProt: P16112 # Defects in AGC1 are the cause of spondyloepiphyseal dysplasia type Kimberley (SEDK) [MIM:608361]. Spondyloepiphyseal dysplasias are a heterogeneous group of congenital chondrodysplasias that specifically affect epiphyses and vertebrae. The autosomal dominant SEDK is associated with premature degenerative arthropathy.
SIMILARITY: Belongs to the aggrecan/versican proteoglycan family. & Contains 1 C-type lectin domain. & Contains 1 EGF-like domain. & Contains 1 Ig-like V-type (immunoglobulin-like) domain. & Contains 4 Link domains. & Contains 1 Sushi (CCP/SCR) domain.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
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 ConditionsMT5-MMP is stable until the expiry date given on the label of stored at -70°C. The protein can be kept at -20°C for several weeks and on ice for several days. Repeated freezing and thawing should be avoided.
Packaging Information
Material Size50 µg
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Katalógusszám GTIN
CC1890 04053252401374

Documentation

Aggrecan Interglobular Domain, Recombinant, His-tagged MSDS

Title

Safety Data Sheet (SDS) 

Aggrecan Interglobular Domain, Recombinant, His-tagged Certificates of Analysis

TitleLot Number
AGGRECAN RECOMBINANT INTERGLOBULAR DOMAIN T331 #8211; G458 (Aggrecan-IGD1) - 2115906 2115906
AGGRECAN RECOMBINANT INTERGLOBULAR DOMAIN T331 – G458 (Aggrecan-IGD1) 3076523
AGGRECAN -2632547 2632547
AGGRECAN -2689130 2689130
AGGRECAN -2721488 2721488
AGGRECAN -2822064 2822064
AGGRECAN RECOMBINANT INTERGLOBULAR DOMAIN T331 #8211; G458 (Aggrecan-IGD1) - 1644072B 1644072B
AGGRECAN RECOMBINANT INTERGLOBULAR DOMAIN T331 – G458 (Aggrecan-IGD1) - 2517318 2517318
AGGRECAN RECOMBINANT INTERGLOBULAR DOMAIN T331 – G458 (Aggrecan-IGD1) - 3416095 3416095
AGGRECAN RECOMBINANT INTERGLOBULAR DOMAIN T331 – G458 (Aggrecan-IGD1) - 3442831 3442831

References

Reference overviewPub Med ID
Cartilage proteoglycans.
Knudson, C B and Knudson, W
Semin. Cell Dev. Biol., 12: 69-78 (2001)  2001

Kivonat megmutatása
11292372 11292372

Data Sheet

Title
AGGRECAN RECOMBINANT INTERGLOBULAR DOMAIN T331 – G458 (Aggrecan-IGD1)