Aggrecanase-I (ADAM-TS4) Enzyme Class and Function.

A disintegrin and metalloproteinase with thrombospondin motifs 4 is an enzyme that in humans is encoded by the ADAMTS4 gene. This gene encodes a member of the ADAM-TS (A Disintegrin And Metalloproteinase with thrombospondin motifs) protein family. 1 Members of the family share several distinct protein modules, including a propeptide region, a metalloproteinase domain, a disintegrin-like domain, and a Thrombo-Spondin 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. 1 Aggrecan degradation, an important factor in the erosion of articular cartilage in arthritic diseases, involves proteolysis of the aggrecan core protein near the N terminus, where 2 major cleavage sites have been identified. Matrix metalloproteinases (MMPs) cleave aggrecan between asn341 and phe342. Tortorella et al. found that human aggrecanase-1 cleaved bovine aggrecan between the glu373-ala374, but not the asn341-phe342, bond.2 By RT-PCR, Tortorella observed upregulation of the aggrecanase-1 message in stimulated human fetal chondrocytes and in joint tissues from adjuvant arthritic rats. Pratta et al. studied the production of ADAM-TS4 in monolayer chondrocytes, capsular fibroblasts, and cartilage.3 They concluded that ADAM-TS4 is constitutively produced in these cells and this tissue, and that stimulation by interleukin-1 results in aggrecanase activation. Thus, the activator could be a potential target by which to control aggrecanase-mediated degradation in arthritic diseases. Structure.

Figure I.

Figure II.

1 2 3

Journal of Biochemistry, (2005) 386, pp.1527 The Journal of Biological Chemistry, (2008), Vol 284, No. 36, pp. 24485-24191

Figure III. Crystal structure of ADMATS-4 and ADAMTS-5 with inhibitors. Illustration of the interactions between (a) marimastat, (b) molecule 8, and (c) molecule 11 with the active site of the catalytic domain of ADAMTS-5.2,4

ADAMTS-4, is the best characterized of the aggrecanases, C-terminal processing of the 75 kDa full-length active form produces isoforms of 60 and 50 kDa, and this results in release of the enzyme from the ECM and alteration of its activity profile. Aggrecan is cleaved at several sites by both MMPs and aggrecanases, however, the signature aggrecanase cleavage site is at Glu373Ala374. The full-length 75 kDa ADAMTS-4 is the most potent aggrecanase of the three isoforms, but it cleaves at Glu1480Gly1481. In contrast, cleavage at Glu373Ala374 is a property of the 60 and 50 kDa isoforms and recombinant truncated proteins lacking the C-terminal spacer region. Thus the C-terminal spacer region can act to inhibit the Glu373 Ala374 aggrecanase activity of full-length ADAMTS-4.4 Active Site Features.

Figure IV.

Protein Science (2008), 17, pp. 1621.

Figure V. Endogenous Substrates

Mechanism of Catalysis