Regulation of ADAMTS9 Secretion and Enzymatic Activity by Its Propeptide*

By B. Koo, Jean-Michel Longpré, R. Somerville, J. Alexander, R. Leduc, S. Apte. 2007

66 citations recorded by the source corpus.

ADAMTS9 is a secreted, cell-surface-binding metalloprotease that cleaves the proteoglycans versican and aggrecan. Unlike most precursor proteins, the ADAMTS9 zymogen (pro-ADAMTS9) is resistant to intracellular processing. Instead, pro-ADAMTS9 is processed by furin at the cell surface. Here, we investigated the role of the ADAMTS9 propeptide in regulating its secretion and proteolytic activity. Removal of the propeptide abrogated secretion of the ADAMTS9 catalytic domain, and secretion was inefficiently restored by expression of the propeptide in trans. Substitution of Ala for Asn residues within each of three consensus N-linked glycosylation sites in the propeptide abrogated ADAMTS9 secretion. Thus, the propeptide is an intramolecular chaperone whose glycosylation is critical for secretion of the mature enzyme. In addition to two previously identified furin-processing sites (Arg74↓ and Arg287↓) the ADAMTS9 propeptide was also furin-processed at Arg209. Substitution of Ala for Arg74, Arg209, and Arg287 resulted in secretion of an unprocessed zymogen. Unexpectedly, versican incubated with cells expressing this pro-ADAMTS9 was processed to a greater extent than when incubated with cells expressing wild-type, furin-processable ADAMTS9. Moreover, cells and medium treated with the proprotein convertase inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethyl ketone had greater versican-cleaving activity than untreated cells. Following furin processing of pro-ADAMTS9, propeptide fragments maintained a non-covalent association with the catalytic domain. Collectively, these observations suggest that, unlike other metalloproteases, furin processing of the ADAMTS9 propeptide reduces its catalytic activity. Thus, the propeptide is a key functional domain of ADAMTS9, mediating an unusual regulatory mechanism that may have evolved to ensure maximal activity of this protease at the cell surface.

Subfields: Immunology and Allergy, Molecular Biology, Oncology

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Immunology and Allergy

1. S. Apte
2. S. Zhong
3. R. Khalil
4. T. Mead

In Molecular Biology

1. S. Apte
2. R. Khalil
3. S. Zhong
4. Carlos Fernandez-Patron
5. D. Edwards
6. Pu Zhang
7. Mengcheng Shen
8. Z. Kassiri

In Oncology

1. S. Apte
2. S. Zhong
3. Carlos Fernandez-Patron
4. D. Edwards
5. Pu Zhang
6. R. Khalil
7. Mengcheng Shen
8. Z. Kassiri

Citations in the corpus

• A Disintegrin and Metalloproteinase (ADAM) and ADAM with Thrombospondin Motifs (ADAMTS) Family in Vascular Biology and Disease (2019, background): “ADAMTS-13 has an unusually short propeptide, and ADAMTS-9 and ADAMTS-13 zymogens appear to be active despite retention of the propeptide [39, 40].”
• The ADAMTS (A Disintegrin and Metalloproteinase with Thrombospondin motifs) family (2015, background): “Other ADAMTS precursors (such as proADAMTS5) are activated by furin not in the Golgi but extracellularly [20], whereas pro-ADAMTS9 is activated on the cell surface when in a complex with the chaperone heat shock protein gp96/GRP94 [21].”
• ADAMTS proteins in human disorders (2018, background): “Koo BH, Longpre JM, Somerville RP, Alexander JP, Leduc R, Apte SS. Regulation of ADAMTS9 secretion and enzymatic activity by its propeptide.”
• ADAMTS9 is a cell-autonomously acting, anti-angiogenic metalloprotease expressed by microvascular endothelial cells. (2010, methodology): “Mammalian expression plasmids for expression of full-length ADAMTS9 or catalytically inactive mutants (Glu(435)Gln or Glu(435)Ala), for C-terminally truncated ADAMTS9 extending up to TSR 8 (designated ADAMTS9 N-L2), or its mutants containing substitution of Ala for Arg (ArgAla) or Ala for Glu(435) (”
• ADAMs family and relatives in cardiovascular physiology and pathology. (2016, background): “ProADAM-TS5 is activated by furin extracellularly [111], whereas proADAM-TS9 is activated on the cell surface when in a complex with the chaperone heart shock protein gp96/GRP94 [112].”
• Biosynthesis and Expression of a Disintegrin-like and Metalloproteinase Domain with Thrombospondin-1 Repeats-15 (2013, methodology): “6B) in awell established assay (6, 12, 17, 29), full-length (ADAMTS15-I) (Fig.”