Structural Basis for Regulated Proteolysis by the α-Secretase ADAM10

Seegar, T.C.M.; Killingsworth, Lauren; Saha, Nayanendu; Meyer, Peter; Patra, Dhabaleswar; Zimmerman, Brandon; Janes, Peter W.; Rubinstein, Eric; Nikolov, Dimitar B.; Skiniotis, Georgios; Kruse, Andrew C.; Blacklow, Stephen C.

doi:10.1016/j.cell.2017.11.014

Cited by 128 publications

(156 citation statements)

References 60 publications

(75 reference statements)

Supporting

Mentioning

135

Contrasting

Order By: Relevance

“…Whereas the first cleavage is assumed to happen in an intramolecular reaction as discussed above, the second cleavage, which led to the generation of the 50-kDa fragment, may alternatively occur in an intermolecular reaction. This possibility is in line with the recently determined crystal structure of the recombinant mADAM10 ectodomain (33), which corresponds to our 50-kDa fragment. Of interest, in the crystal structure, mADAM10 was observed as a tetramer, where the Cterminal amino acids of one mADAM10 molecule were bound in the active site of a neighboring mADAM10 molecule, which suggests that an intermolecular cleavage at this mADAM10 site, in principle, may be possible.…”

Section: Discussionsupporting

confidence: 91%

“…After signal peptide cleavage in the endoplasmic reticulum, the proteolytically inactive, immature ADAM (proA-DAM10) transits via the secretory pathway, where it is cleaved by furin or homologous proteases and converted to the proteolytically active, mature ADAM10 (mADAM10), which cleaves its substrates at or close to the cell surface. A recently reported crystal structure shows that the mature ectodomain adopts a compact, arrowhead-like structure in which the metalloprotease domain is enveloped by the disintegrin and cysteine-rich domains, the latter of which partially blocks the active site (33).…”

mentioning

confidence: 99%

See 1 more Smart Citation

The metalloprotease ADAM10 (a disintegrin and metalloprotease 10) undergoes rapid, postlysis autocatalytic degradation

et al. 2018

View full text Add to dashboard Cite

The transmembrane protein, ADAM10 (a disintegrin and metalloprotease 10), has key physiologic functions-for example, during embryonic development and in the brain. During transit through the secretory pathway, immature ADAM10 (proADAM10) is converted into its proteolytically active, mature form (mADAM10). Increasing or decreasing the abundance and/or activity of mADAM10 is considered to be a therapeutic approach for the treatment of such diseases as Alzheimer's disease and cancer. Yet biochemical detection and characterization of mADAM10 has been difficult. In contrast, proADAM10 is readily detected-for example, in immunoblots-which suggests that mADAM10 is only a fraction of total cellular ADAM10. Here, we demonstrate that mADAM10, but not proADAM10, unexpectedly undergoes rapid, time-dependent degradation upon biochemical cell lysis in different cell lines and in primary neurons, which prevents the detection of the majority of mADAM10 in immunoblots. This degradation required the catalytic activity of ADAM10, was efficiently prevented by adding active site inhibitors to the lysis buffer, and did not affect proADAM10, which suggests that ADAM10 degradation occurred in an intramolecular and autoproteolytic manner. Inhibition of postlysis autoproteolysis demonstrated efficient cellular ADAM10 maturation with higher levels of mADAM10 than proADAM10. Moreover, a cycloheximide chase experiment revealed that mADAM10 is a long-lived protein with a half-life of approximately 12 h. In summary, our study demonstrates that mADAM10 autoproteolysis must be blocked to allow for the proper detection of mADAM10, which is essential for the correct interpretation of biochemical and cellular studies of ADAM10.-Brummer, T., Pigoni, M., Rossello, A., Wang, H., Noy, P. J., Tomlinson, M. G., Blobel, C. P., Lichtenthaler, S. F. The metalloprotease ADAM10 (a disintegrin and metalloprotease 10) undergoes rapid, postlysis autocatalytic degradation.

show abstract

Section: Discussionsupporting

confidence: 91%

mentioning

confidence: 99%

The metalloprotease ADAM10 (a disintegrin and metalloprotease 10) undergoes rapid, postlysis autocatalytic degradation

et al. 2018

View full text Add to dashboard Cite

show abstract

“…Sheddases recognize amino acid motifs and/or secondary structures in their substrates Sheddases often have preferences for certain amino acid motifs, which is seen in in vitro assays (e.g., Gruninger-Leitch et al, 2002;Caescu et al, 2009), by mutational analyses (e.g., Sisodia, 1992) and by sheddase structure determinations (e.g., Hong et al, 2000;Seegar et al, 2017). However, this requirement is less pronounced compared to many soluble proteases, such as trypsin and caspases.…”

Section: Substrate Recognition Of Sheddasesmentioning

confidence: 99%

Proteolytic ectodomain shedding of membrane proteins in mammals—hardware, concepts, and recent developments

2018

View full text Add to dashboard Cite

Proteolytic removal of membrane protein ectodomains (ectodomain shedding) is a post-translational modification that controls levels and function of hundreds of membrane proteins. The contributing proteases, referred to as sheddases, act as important molecular switches in processes ranging from signaling to cell adhesion. When deregulated, ectodomain shedding is linked to pathologies such as inflammation and Alzheimer's disease. While proteases of the "a disintegrin and metalloprotease" (ADAM) and "beta-site APP cleaving enzyme" (BACE) families are widely considered as sheddases, in recent years a much broader range of proteases, including intramembrane and soluble proteases, were shown to catalyze similar cleavage reactions. This review demonstrates that shedding is a fundamental process in cell biology and discusses the current understanding of sheddases and their substrates, molecular mechanisms and cellular localizations, as well as physiological functions of protein ectodomain shedding. Moreover, we provide an operational definition of shedding and highlight recent conceptual advances in the field. While new developments in proteomics facilitate substrate discovery, we expect that shedding is not a rare exception, but rather the rule for many membrane proteins, and that many more interesting shedding functions await discovery.

show abstract

“…Because no structural information for any isolated ADAM prodomain nor of any pro‐ADAM protease is available, we used secondary structure prediction tools and HHpred (37, 38) for remote homology modeling and 3D structure prediction for ADAM prodomains. We used the already known structures of the ADAM10 (43) and ADAM22 (44) ectodomains to obtain structural insight in the postulated activation mechanism. In accordance with a previous report (27), our analysis characterized the prodomains of ADAM9, 10, and 17 to have a central folded core region, which is C‐terminally flanked by an unstructured linker region.…”

Section: Resultsmentioning

confidence: 99%

Meprin β induces activities of A disintegrin and metalloproteinases 9, 10, and 17 by specific prodomain cleavage

Wichert¹,

Scharfenberg²,

Colmorgen³

et al. 2019

The FASEB Journal

View full text Add to dashboard Cite

Meprin β is a membrane‐bound metalloprotease involved in extracellular matrix assembly and inflammatory processes in health and disease. A disintegrin and metalloproteinase (ADAM)10 and ADAM17 are physiologic relevant sheddases of inactive promeprin β, which influences its substrate repertoire and subsequent biologic functions. Proteomic analysis also revealed several ADAMs as putative meprin β substrates. Here, we demonstrate specific N‐terminal processing of ADAM9, 10, and 17 by meprin β and identify cleavage sites within their prodomains. Because ADAM prodomains can act as specific inhibitors, we postulate a role for meprin β in the regulation of ADAM activities. Indeed, prodomain cleavage by meprin β caused increased ADAM protease activities, as observed by peptide‐based cleavage assays and demonstrated by increased ectodomain shedding activity. Direct interaction of meprin β and ADAM proteases could be shown by immunofluorescence microscopy and immunoprecipitation experiments. As demonstrated by a bacterial activator of meprin β and additional measurement of TNF‐α shedding on bone marrow‐derived macrophages, meprin β/ADAM protease interactions likely influence inflammatory conditions. Thus, we identified a novel proteolytic pathway of meprin β with ADAM proteases to control protease activities at the cell surface as part of the protease web.—Wichert, R., Scharfenberg, F., Colmorgen, C., Koudelka, T., Schwarz, J., Wetzel, S., Potempa, B., Potempa, J., Bartsch, J. W., Sagi, I., Tholey, A., Saftig, P., Rose‐John, S., Becker‐Pauly, C. Meprin β induces activities of A disintegrin and metalloproteinases 9, 10, and 17 by specific prodomain cleavage. FASEB J. 33, 11925‐11940 (2019). http://www.fasebj.org

show abstract

Structural Basis for Regulated Proteolysis by the α-Secretase ADAM10

Cited by 128 publications

References 60 publications

The metalloprotease ADAM10 (a disintegrin and metalloprotease 10) undergoes rapid, postlysis autocatalytic degradation

The metalloprotease ADAM10 (a disintegrin and metalloprotease 10) undergoes rapid, postlysis autocatalytic degradation

Proteolytic ectodomain shedding of membrane proteins in mammals—hardware, concepts, and recent developments

Meprin β induces activities of A disintegrin and metalloproteinases 9, 10, and 17 by specific prodomain cleavage

Contact Info

Product

Resources

About