title: Selenoprotein K is a novel target of m-calpain, and cleavage is regulated by Toll-like receptor-induced calpastatin in macrophages.
abstract: Calpains are proteolytic enzymes that modulate cellular function through cleavage of targets, thereby modifying their actions. An important role is emerging for calpains in regulating inflammation and immune responses, although specific mechanisms by which this occurs have not been clearly defined. In this study, we identify a novel target of calpain, selenoprotein K (SelK), which is an endoplasmic reticulum transmembrane protein important for Ca(2+) flux in immune cells. Calpain-mediated cleavage of SelK was detected in myeloid cells (macrophages, neutrophils, and dendritic cells) but not in lymphoid cells (B and T cells). Both m- and μ-calpain were capable of cleaving immunoprecipitated SelK, but m-calpain was the predominant isoform expressed in mouse immune cells. Consistent with these results, specific inhibitors were used to show that only m-calpain cleaved SelK in macrophages. The cleavage site in SelK was identified between Arg(81) and Gly(82) and the resulting truncated SelK was shown to lack selenocysteine, the amino acid that defines selenoproteins. Resting macrophages predominantly expressed cleaved SelK and, when activated through different Toll-like receptors (TLRs), SelK cleavage was inhibited. We found that decreased calpain cleavage was due to TLR-induced up-regulation of the endogenous inhibitor, calpastatin. TLR-induced calpastatin expression not only inhibited SelK cleavage, but cleavage of another calpain target, talin. Moreover, the expression of the calpain isoforms and calpastatin in macrophages were different from T and B cells. Overall, our findings identify SelK as a novel calpain target and reveal dynamic changes in the calpain/calpastatin system during TLR-induced activation of macrophages.
authors.affiliation: Department of Cell and Molecular Biology, John A Burns School of Medicine, University of Hawaii, Honolulu, Hawaii 96813, USA.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Huang
authors.forename: Fukun W
authors.initials: FW
authors.lastname: Hoffmann
authors.forename: Robert L
authors.initials: RL
authors.lastname: Norton
authors.forename: Ann C
authors.initials: AC
authors.lastname: Hashimoto
authors.forename: Peter R
authors.initials: PR
authors.lastname: Hoffmann
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002154:Calpain / Q000378:metabolism*; D002465:Cell Movement; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D051140:Selenoproteins / Q000378:metabolism*; D051193:Toll-Like Receptors / Q000378:metabolism*
chemical_list: D002135:Calcium-Binding Proteins; D008024:Ligands; C495239:Selenof protein, mouse; D051140:Selenoproteins; D051193:Toll-Like Receptors; C033668:calpastatin; D002154:Calpain; C449000:m-calpain; D002118:Calcium