English:
[0002] ADAM ("a -Λsintegrin and metalloproteinase") proteins represent a family of membrane-associated multidomain zinc-dependent metalloproteases with high sequence homology and domain organization. In addition to the disintegrin and protease domains, the ADAM proteins generally contain a prodomain, a cysteine-rich domain, an EGF-like domain, a transmembrane domain, and a cytoplasmic tail domain. The ADAM proteins are unique among cell-surface proteins in containing features of both adhesive proteins and proteases (Kaushal and Shah, J. Clin. Invest. 105:1335 (2000)).
[0003] Recently, new members of the ADAM family have been identified which lack the transmembrane and cytoplasmic tail domains. More importantly, these new members contain unique thrombospondin type I repeats (TSRs) not found in other ADAMs. These ADAMTS ("α -Λsintegrin and metalloproteinase with thrombospondin motifs") proteins also contain a prodomain, a metalloprotease domain, a disintegrin domain, a cysteine-rich domain, and a spacer region, and may also contain a PLAC domain which is a 30-40 amino acid peptide containing six cysteins. Like other ADAM proteins, all ADAMTS proteins identified to date contain the catalytic consensus sequence HXYG-ΩΗD, which coordinates the Zn2+ion necessary for protease activity (Tang, Int. J. Biochem. Cell Biol. 445:223 (2001)).
[0004] The members of the ADAMTS family, which now number over twenty, all contain a single TSR following the disintegrin domain; this internal TSR has been shown to bind to heparin (Kuno et al., J. Biol. Chem. 272:556 (1997)). The ADAMTSs, however, can be distinguished from each other, in part, by the variable number of C-terminal TSRs they contain downstream of the spacer region. For example, ADAMTS-4 contains no C-terminal TSRs, ADAMTS-5 contains one C- terminal TSR, ADAMTS- 1 (as well as the human homolog METH1) and ADAMTS- 16 contain two C-terminal TSRs, ADAMTS- 10 and -18 contain five C-terminal TSRs, and ADAMTS-9 and -20 contain fourteen C-terminal TSRs.
[0005] The ADAMTSs have been implicated in a variety of pathological disorders. For example, mutations in ADAMTS-2 result in Ehlers-Danlos syndrome in humans and dermatosparaxis in cattle (Colige et al., Am. J. Hum. Genet. 65:308 (1999)), while mutations in ADAMTS- 13 (also known as the von Willebrand factor cleaving protein) result in thrombotic thrombocytopenic purpura (Kokame et al., Proc. Natl. Acad. Sci. USA 99:11902 (2002)).
[0007] The ADAMTS aggrecanases cleave between amino acids Glu373-Ala374within the interglobular domain of the GI globular domain of aggrecan, which exposes an N- terminal neoepitope (374ARGSV) on the resulting C-terminal aggrecan fragment (Tortorella et al., Matrix Biol. 21 :499 (2002); Westling et al., J. Biol. Chem. 277:16059 (2002); Tortorella et al., J. Biol. Chem. 275:18566 (2000)). This ARGSV aggrecan fragment has been found in synovial fluid from patients with inflammatory joint disease, joint injury, and OA (Malfait et al., J. Biol. Chem. 277:22201 (2002); Lohmander et al, Arthritis Rheum. 36:1214 (1993); Sandy et al., J. Clin. Invest. 89:1512 (1992)). In addition, the resulting N-terminal aggrecan fragment containing the C-terminal NITEGE373neoepitope has been found in articular cartilage from patients with joint injury, OA, and RA (Malfait et al., supra; Sandy and Verscharen, Biochem. J. 358:615 (2001); Lark et al., J. Clin. Invest. 100:93 (1997)). Inhibition of aggrecanase activity with a synthetic ADAMTS inhibitor has been shown to prevent aggrecan degradation in osteoarthritic cartilage, as measured by release of aggrecan fragments containing the374ARGSV neoepitope (Malfait et al., supra).
[0009] United States Patent Application Publication No. 2004/0044194 Al , incorporated herein in its entirety by reference, relates to ADAMTS 18 nucleic acid molecules and polypeptides encoded thereby.
[0010] United States Patent Application Publication No. 2004/0054149 Al, incorporated herein in its entirety by reference, relates to truncated ADAMTS molecules and preferably truncated ADAMTS-4 (aggrecanase- 1) and ADAMTS-5 (aggrecanase-2) nucleic acid molecules and polypeptides encoded thereby. Simularly, U.S. Patent Application Publication No. 2004/0142863 Al, incorporated herein in its entirety by reference, relates to truncated ADAMTS-4 nucleic acid molecules and polypeptides encoded thereby.

中文:
[0002] ADAM(“解整连蛋白(disintegrin)和金属蛋白酶”)蛋白代表具有高度序列同源性和结构域组织的膜相关多结构域锌依赖性金属蛋白酶家族。除了解整连蛋白和蛋白酶结构域外,ADAM蛋白通常含有前结构域(predomain)、富含半胱氨酸的结构域、EGF样结构域、跨膜结构域以及胞质尾结构域。ADAM蛋白是细胞表面蛋白中独特的,这在于其含有粘着蛋白和蛋白酶双特性(Kaushal和Shah,J.Clin.Invest.105:1335(2000))。
[0003]最近,已鉴定了缺乏跨膜和胞质尾结构域的ADAM家族的新成员。更重要的是,这些新成员含有未在其他ADAM中发现的独特的血小板反应蛋白I型重复(TSRs)。这些ADAMTS(“具有血小板反应蛋白基序的解整连蛋白和金属蛋白酶”)蛋白也含有前结构域、金属蛋白酶结构域、解整连蛋白结构域、富含半胱氨酸的结构域和间隔区,并且可能也含有PLAC结构域,其是含有6个半胱氨酸的30-40个氨基酸的肽。如同其它ADAM蛋白,所有迄今鉴定的ADAMTS蛋白都含有催化性共有序列HXXGXXHD,其协调蛋白酶活性必需的Zn2+离子(Tang,Int.J.Biochem.Cell Biol. 445:223(2001))。
[0004]ADAMTS家族的成员,其现在数目超过20个,所有都在解整连蛋白结构域后含有一个TSR;已表明此内部TSR结合肝素(Kuno等,J,Biol.Chem.272:556(1997))。然而,ADAMTS相互之间可以区分,这部分地通过它们在间隔区下游所含有的可变数目的C-末端TSR实现。例如,ADAMTS-4不含有C-末端TSR,ADAMTS-5含有1个C-末端TSR,ADAMTS-1(以及人同源物METH1)和ADAMTS-16含有2个C-末端TSR,ADAMTS-10和-18含有5个C-末端TSR,以及ADAMTS-9和-20含有14个C-末端TSR。
[0005]ADAMTS已牵涉在多种病理障碍中。例如,ADAMTS-2的突变导致人类Ehlers-Danlos综合征和牛的皮肤脆裂症(Colige等,Am.J.Hum.Genet.65:308(1999)),而ADAMTS-13(也称作vonWillebrand因子切割蛋白)的突变导致血栓形成性血小板减少性紫癜(Kokame等,Proc.Natl.Acad.Sci.USA 99:11902(2002))。
[0007]ADAMTS软骨聚集蛋白聚糖酶在软骨聚集蛋白聚糖的G1球形结构域的球间结构域内的氨基酸Glu373-Ala374之间切割,其暴露所得C-末端软骨聚集蛋白聚糖片段上的N-末端的新表位(neoepitope)(374ARGSV)(Tortorella等,Matrix Biol.21:499(2002);Westling等,J.Biol.Chem.277:16059(2002);Tortorella等,J.Biol.Chem.275:18566(2000))。已在患有炎性关节病、关节损伤和OA患者的滑液中发现了该374ARGSV软骨聚集蛋白聚糖片段(Malfait等,J.Biol.Chem.277:22201(2002);Lohmander等,ArthritisRheum.36:1214(1993);Sandy等,J.Clin.Invest.89:1512(1992))。另外,已在患有关节损伤、OA和RA患者的关节软骨中发现了所得的含有C-末端NITEGE373新表位的N-末端软骨聚集蛋白聚糖片段(Malfait等,见上;Sandy和Verscharen,Biochem.J.358:615(2001);Lark等,J.Clin.Invest.100:93(1997))。已表明用合成的ADAMTS抑制剂抑制软骨聚集蛋白聚糖酶活性能在骨关节炎软骨中阻止软骨聚集蛋白聚糖降解,如通过含有374ARGSV新表位的软骨聚集蛋白聚糖片段的释放所测量的(Malfait等,见上)。
[0009]美国专利申请公开No.2004/0044194 A1,在此以其整体引入作为参考,涉及ADAMTS 18核酸分子和其编码的多肽。
[0010]美国专利申请公开No.2004/0054149 A1,在此以其整体引入作为参考,涉及截短的ADAMTS分子,且优选地为截短的ADAMTS-4(软骨聚集蛋白聚糖酶-1)和ADAMTS-5(软骨聚集蛋白聚糖酶-2)核酸分子及其编码的多肽。类似地,美国专利申请公开No.2004/0142863 A1,在此以其整体引入作为参考,涉及截短的ADAMTS-4核酸分子及其编码的多肽。