Aggrecan-IGD1

Description
Catalogue Number	CC1890
Brand Family	Chemicon®
Trade Name	Chemicon
Description	Aggrecan Interglobular Domain, Recombinant, His-tagged
Overview	Molecular 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 Information	Aggrecan 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].

Description

Catalogue Number

CC1890

Brand Family

Chemicon®

Trade Name

Chemicon

Description

Aggrecan Interglobular Domain, Recombinant, His-tagged

Overview

Molecular 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 Information

Aggrecan 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].

Product Information
Presentation	The 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.

Product Information

Presentation

The 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.

Applications
Key Applications	Affects Function

Applications

Key Applications

Affects Function

Biological Information
Source	Human
Entrez Gene Number	NM_001135.2 NM_013227.2
Entrez Gene Summary	This 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	P16112
UniProt Summary	FUNCTION: 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.

Biological Information

Source

Human

Entrez Gene Number

Entrez Gene Summary

This 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

P16112

UniProt Summary

FUNCTION: 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.

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	MT5-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.

Storage and Shipping Information

Storage Conditions

MT5-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 Size	50 µg

Packaging Information

Material Size

50 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

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