Quantitative Proteomics Screen Identifies a Substrate Repertoire of Rhomboid Protease RHBDL2 in Human Cells and Implicates It in Epithelial Homeostasis

Overview

Journal Sci Rep

Specialty Science

Date 2017 Aug 6

PMID 28779096

Citations 18

Authors

Nicholas Johnson

Jana Brezinova

Elaine Stephens

Emma Burbridge

Matthew Freeman

Colin Adrain

Kvido Strisovsky

Affiliations

Soon will be listed here.

Abstract

Rhomboids are intramembrane serine proteases conserved in all kingdoms of life. They regulate epidermal growth factor receptor signalling in Drosophila by releasing signalling ligands from their transmembrane tethers. Their functions in mammals are poorly understood, in part because of the lack of endogenous substrates identified thus far. We used a quantitative proteomics approach to investigate the substrate repertoire of rhomboid protease RHBDL2 in human cells. We reveal a range of novel substrates that are specifically cleaved by RHBDL2, including the interleukin-6 receptor (IL6R), cell surface protease inhibitor Spint-1, the collagen receptor tyrosine kinase DDR1, N-Cadherin, CLCP1/DCBLD2, KIRREL, BCAM and others. We further demonstrate that these substrates can be shed by endogenously expressed RHBDL2 and that a subset of them is resistant to shedding by cell surface metalloproteases. The expression profiles and identity of the substrates implicate RHBDL2 in physiological or pathological processes affecting epithelial homeostasis.

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References

Ribatti D, Crivellato E . "Sprouting angiogenesis", a reappraisal. Dev Biol. 2012; 372(2):157-65. DOI: 10.1016/j.ydbio.2012.09.018. View

Lemberg M, Freeman M . Functional and evolutionary implications of enhanced genomic analysis of rhomboid intramembrane proteases. Genome Res. 2007; 17(11):1634-46. PMC: 2045146. DOI: 10.1101/gr.6425307. View

Zhong X, Rescorla F . Cell surface adhesion molecules and adhesion-initiated signaling: understanding of anoikis resistance mechanisms and therapeutic opportunities. Cell Signal. 2011; 24(2):393-401. DOI: 10.1016/j.cellsig.2011.10.005. View

Phin S, Marchand-Adam S, Fabre A, Marchal-Somme J, Bantsimba-Malanda C, Kataoka H . Imbalance in the pro-hepatocyte growth factor activation system in bleomycin-induced lung fibrosis in mice. Am J Respir Cell Mol Biol. 2009; 42(3):286-93. DOI: 10.1165/rcmb.2008-0305OC. View

Wunderle L, Knopf J, Kuhnle N, Morle A, Hehn B, Adrain C . Rhomboid intramembrane protease RHBDL4 triggers ER-export and non-canonical secretion of membrane-anchored TGFα. Sci Rep. 2016; 6:27342. PMC: 4893610. DOI: 10.1038/srep27342. View

Cheng T, Lai C, Jiang S, Hung J, Liu S, Chang B . RHBDL2 is a critical membrane protease for anoikis resistance in human malignant epithelial cells. ScientificWorldJournal. 2014; 2014:902987. PMC: 4058132. DOI: 10.1155/2014/902987. View

Logue S, Cleary P, Saveljeva S, Samali A . New directions in ER stress-induced cell death. Apoptosis. 2013; 18(5):537-46. DOI: 10.1007/s10495-013-0818-6. View

Cheng T, Wu Y, Lin H, Hsu F, Liu S, Chang B . Functions of rhomboid family protease RHBDL2 and thrombomodulin in wound healing. J Invest Dermatol. 2011; 131(12):2486-94. DOI: 10.1038/jid.2011.230. View

Leitinger B . Transmembrane collagen receptors. Annu Rev Cell Dev Biol. 2011; 27:265-90. DOI: 10.1146/annurev-cellbio-092910-154013. View

10.

Nilse L, Avci D, Heisterkamp P, Serang O, Lemberg M, Schilling O . Yeast membrane proteomics using leucine metabolic labelling: Bioinformatic data processing and exemplary application to the ER-intramembrane protease Ypf1. Biochim Biophys Acta. 2016; 1864(10):1363-71. DOI: 10.1016/j.bbapap.2016.07.002. View

11.

Kall L, Krogh A, Sonnhammer E . A combined transmembrane topology and signal peptide prediction method. J Mol Biol. 2004; 338(5):1027-36. DOI: 10.1016/j.jmb.2004.03.016. View

12.

Adrain C, Strisovsky K, Zettl M, Hu L, Lemberg M, Freeman M . Mammalian EGF receptor activation by the rhomboid protease RHBDL2. EMBO Rep. 2011; 12(5):421-7. PMC: 3090019. DOI: 10.1038/embor.2011.50. View

13.

Noy P, Swain R, Khan K, Lodhia P, Bicknell R . Sprouting angiogenesis is regulated by shedding of the C-type lectin family 14, member A (CLEC14A) ectodomain, catalyzed by rhomboid-like 2 protein (RHBDL2). FASEB J. 2016; 30(6):2311-23. DOI: 10.1096/fj.201500122R. View

14.

Hruz T, Laule O, Szabo G, Wessendorp F, Bleuler S, Oertle L . Genevestigator v3: a reference expression database for the meta-analysis of transcriptomes. Adv Bioinformatics. 2009; 2008:420747. PMC: 2777001. DOI: 10.1155/2008/420747. View

15.

Vizcaino J, Cote R, Csordas A, Dianes J, Fabregat A, Foster J . The PRoteomics IDEntifications (PRIDE) database and associated tools: status in 2013. Nucleic Acids Res. 2012; 41(Database issue):D1063-9. PMC: 3531176. DOI: 10.1093/nar/gks1262. View

16.

Sahin U, Weskamp G, Kelly K, Zhou H, Higashiyama S, Peschon J . Distinct roles for ADAM10 and ADAM17 in ectodomain shedding of six EGFR ligands. J Cell Biol. 2004; 164(5):769-79. PMC: 2172154. DOI: 10.1083/jcb.200307137. View

17.

Junttila M, Saarinen S, Schmidt T, Kast J, Westermarck J . Single-step Strep-tag purification for the isolation and identification of protein complexes from mammalian cells. Proteomics. 2005; 5(5):1199-203. DOI: 10.1002/pmic.200400991. View

18.

Higy M, Junne T, Spiess M . Topogenesis of membrane proteins at the endoplasmic reticulum. Biochemistry. 2004; 43(40):12716-22. DOI: 10.1021/bi048368m. View

19.

Brambrink T, Foreman R, Welstead G, Lengner C, Wernig M, Suh H . Sequential expression of pluripotency markers during direct reprogramming of mouse somatic cells. Cell Stem Cell. 2008; 2(2):151-9. PMC: 2276627. DOI: 10.1016/j.stem.2008.01.004. View

20.

Huang H, Chang M, Chen Y, Hsu H, Chiang C, Cheng T . Persistent elevation of hepatocyte growth factor activator inhibitors in cholangiopathies affects liver fibrosis and differentiation. Hepatology. 2011; 55(1):161-72. DOI: 10.1002/hep.24657. View