Phosphorylation of the type II transmembrane serine protease, TMPRSS13, in hepatocyte growth factor activator inhibitor-1 and -2–mediated cell-surface localization

Abstract

TMPRSS13 is a member of the type II transmembrane serine protease (TTSP) family. Although various TTSPs have been characterized in detail biochemically and functionally, the basic properties of TMPRSS13 remain unclear. Here, we investigate the activation, inhibition, post-translational modification, and localization of TMPRSS13. We show that TMPRSS13 is a glycosylated, active protease and that its own proteolytic activity mediates zymogen cleavage. Full-length, active TMPRSS13 exhibits impaired cell-surface expression in the absence of the cognate Kunitz-type serine protease inhibitors, hepatocyte growth factor activator inhibitor (HAI)-1 or HAI-2. Concomitant presence of TMPRSS13 with either HAI-1 or -2 mediates phosphorylation of residues in the intracellular domain of the protease, and it coincides with efficient transport of the protease to the cell surface and its subsequent shedding. Cell-surface labeling experiments indicate that the dominant form of TMPRSS13 on the cell surface is phosphorylated, whereas intracellular TMPRSS13 is predominantly non-phosphorylated. These data provide novel insight into the cellular properties of TMPRSS13 and highlight phosphorylation of TMPRSS13 as a novel post-translational modification of this TTSP family member and potentially other members of this family of proteases.

Establishment of stable Vero cell lines expressing TMPRSS2 and MSPL: A useful tool for propagating porcine epidemic diarrhea virus in the absence of exogenous trypsin

“…Western blot results showed that target bands were examined but the immunoreactive bands are not unique, which is consistent with previous work [26]. We speculated that serine proteases TMPRSS2 and MSPL, the glycosylated proteins, showed multiple cut bands with different sizes by their own proteolytic activity [58]. Moreover, the results indicated that the bands, larger than the target bands (TMPRSS2,82 kDa; MSPL, 89 kDa), were detected in this study, and we speculated that caused by phosphorylation [58].…”

Glycan-Dependent and -Independent Dual Recognition between DC-SIGN and Type II Serine Protease MSPL/TMPRSS13 in Colorectal Cancer Cells

“…Next, to explore the functional aspects of the molecular interaction between DC-SIGN and MSPL/TMPRSS13, we constructed a plasmid containing the MSPL extracellular domain (MSPL-ECD). A stable transfectant of MSPL-ECD showed multiple bands after CBB staining (Figure 3a), as previously reported [24], due to self-cleavage and glycosylation variants. When we conducted the DC-SIGN-ECD lectin blot assay, we found that DC-SIGN-ECD binds to MSPL-ECD in a Ca 2+ -dependent manner (Figure 3b) and that all self-digested peptide fragments contained DC-SIGN-bindable glycans.…”

“…It has been reported that MSPL/TMPRSS13 retains its protease activity when it is phosphorylated, shed, and released from the cell surface [24]. Next, to explore the functional aspects of the molecular interaction between DC-SIGN and MSPL/TMPRSS13, we constructed a plasmid containing the MSPL extracellular domain (MSPL-ECD).…”

“…Given that DC-SIGN binds to MSPL in a Ca 2+ -dependent manner, the results of these co-incubation studies suggest that glycan-mediated MSPL recognition by DC-SIGN interferes with DC-SIGN digestion by MSPL (see Figure S1). phosphorylated, shed, and released from the cell surface [24]. Next, to explore the functional aspects of the molecular interaction between DC-SIGN and MSPL/TMPRSS13, we constructed a plasmid containing the MSPL extracellular domain (MSPL-ECD).…”

Crystal structure of inhibitor-bound human MSPL that can activate high pathogenic avian influenza

“…The refined model contains the hMSPL with the residue range of 188-558, except 319 and 320, decanoyl-RVKR-cmk, and a calcium ion. Glycans attached to residues Asn250 and Asn400 were observed, but no phosphorylated residues found (18).…”