In vivo N-Terminomics Highlights Novel Functions of ADAMTS2 and ADAMTS14 in Skin Collagen Matrix Building

Abstract

A disintegrin and metalloproteinase with thrombospondin type I motif (ADAMTS)2 and ADAMTS14 were originally known for their ability to cleave the aminopropeptides of fibrillar collagens. Previous work using N-terminomic approach (N-TAILS) in vitro led to the identification of new substrates, including some molecules involved in TGF-β signaling. Here, N-TAILS was used to investigate the substrates of these two enzymes in vivo, by comparing the N-terminomes of the skin of wild type mice, mice deficient in ADAMTS2, in ADAMTS14 and in both ADAMTS2 and ADAMTS14. This study identified 68 potential extracellular and cell surface proteins, with the majority of them being cleaved by both enzymes. These analyses comfort their role in collagen matrix organization and suggest their implication in inflammatory processes. Regarding fibrillar collagen, this study demonstrates that both ADAMTS2 and ADAMTS14 are involved in the processing of the aminopropeptide of alpha1 and alpha2 type V collagen. It also revealed the existence of several cleavage sites in the Col1 domain and in the C-propeptide of type I collagens. In addition to collagens and other extracellular proteins, two major components of the cell cytoskeleton, actin and vimentin, were also identified as potential substrates. The latter data were confirmed in vitro using purified enzymes and could potentially indicate other functions for ADAMTS2 and 14. This original investigation of mouse skin degradomes by N-terminomic highlights the essential role of ADAMTS2 and ADAMTS14 in collagen matrix synthesis and turnover, and gives clues to better understand their functions in skin pathophysiology. Data are available via ProteomeXchange with identifier PXD022179.

The Pathogenesis of Cardiac Fibrosis: A Review of Recent Progress

“…Sequential processing steps are required in the extracellular space for procollagen to become a mature fibril collagen. A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) 2, 3, and 14 cleave the N-terminal [ 31 ] (also referred to as procollagen amino-terminal proteinases (PNPs)), whereas bone morphogenic protein (BMP)-1 is the protease responsible for cleaving the C-terminal [ 32 , 33 ] (also referred to as procollagen carboxy-terminal proteinases (PCPs)). Additionally, PCP enhancer (PCPE)-1 and 2, enhancers of BMP-1 activity, facilitate C-terminus cleavage [ 34 ].…”

ADAMTS2 and ADAMTS14 can substitute for ADAMTS3 in adults for pro-VEGFC activation and lymphatic homeostasis

“…ADAMTS3 possesses high similarity with ADAMTS2 and ADAMTS14 in terms of sequence and domain composition. These 3 enzymes also share the capacity to cleave the amino-propeptide of fibrillar collagens but with different efficacy: ADAMTS2 is the most efficient, and ADAMTS3 and ADAMTS14 display only moderate and low activities, respectively (12,14,17,21,22). By sharp contrast, we have shown here that the 3 enzymes display similar processing activities when pro-VEGFC was used as substrate, showing that it might be a more physiological substrate for ADAMTS14 than fibrillar procollagens and, therefore, that ADAMTS14 activity could be more important for lymphangiogenesis than for collagen maturation.…”

Regulation of ADAMTS Proteases

“…Since the primary function of ADAMTS2 is the processing of the N-terminal procollagen propeptide, the functional significance for ADAMTS2 induction in macrophages during wound repair remains to be established. Recently, additional substrates for ADAMTS2 were identified in the skin, including fibronectin, which is an important part of the provisional ECM formed after injury, and several proteins linked to inflammation ( Bekhouche et al, 2016 ; Leduc et al, 2021 ). Therefore, regulation of ADAMTS2 in non-collagen producing cell types could hint to a broader substrate spectrum as previously anticipated in vivo .…”