Tài liệu Báo cáo khoa học: TMPRSS13, a type II transmembrane serine protease, is inhibited by

TMPRSS13, a type II transmembrane serine protease, is

inhibited by hepatocyte growth factor activator inhibitor

type 1 and activates pro-hepatocyte growth factor

Tomio Hashimoto1

, Minoru Kato2

, Takeshi Shimomura2 and Naomi Kitamura1

1 Department of Biological Sciences, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta,

Midori-ku, Yokohama, Japan

2 Advanced Medical Research Laboratory, Mitsubishi Tanabe Pharma Corporation, Kamoshida-cho, Aoba-ku, Yokohama, Japan

Introduction

Type II transmembrane serine proteases (TTSPs) are

structurally defined by the presence of a short N-termi￾nal cytoplasmic domain, a transmembrane domain

located near the N-terminus, and a C-terminal extra￾cellular serine protease domain. In addition, TTSPs

possess a stem region that may contain a diverse array

Keywords

activation of pro-hepatocyte growth factor;

hepatocyte growth factor activator inhibitor

type 1 (HAI-1); Kunitz-type inhibitor;

TMPRSS13; type II transmembrane serine

protease (TTSP)

Correspondence

N. Kitamura, Department of Biological

Sciences, Graduate School of Bioscience

and Biotechnology, Tokyo Institute of

Technology, Nagatsuta, Midori-ku,

Yokohama 226-8501, Japan

Fax: +81 45 924 5771

Tel: +81 45 924 5701

E-mail: [email protected]

(Received 31 May 2010, revised 26 August

2010, accepted 24 September 2010)

doi:10.1111/j.1742-4658.2010.07894.x

Type II transmembrane serine proteases (TTSPs) are structurally defined

by the presence of a transmembrane domain located near the N-terminus

and a C-terminal extracellular serine protease domain. The human TTSP

family consists of 17 members. Some members of the family have pivotal

functions in development and homeostasis, and are involved in tumorigene￾sis and viral infections. The activities of TTSPs are regulated by endoge￾nous protease inhibitors. However, protease inhibitors of most TTSPs have

not yet been identified. In this study, we investigated the inhibitory effect

of hepatocyte growth factor activator inhibitor type 1 (HAI-1), a Kunitz￾type serine protease inhibitor, on several members of the TTSP family. We

found that the protease activity of a member, TMPRSS13, was inhibited

by HAI-1. A detailed analysis revealed that a soluble form of HAI-1 with

one Kunitz domain (NK1) more strongly inhibited TMPRSS13 than

another soluble form of HAI-1 with two Kunitz domains (NK1LK2). In

addition, an in vitro protein binding assay showed that NK1 formed com￾plexes with TMPRSS13, but NK1LK2 did not. TMPRSS13 converted

single-chain pro-hepatocyte growth factor (pro-HGF) to a two-chain form

in vitro, and the pro-HGF converting activity of TMPRSS13 was inhibited

by NK1. The two-chain form of HGF exhibited biological activity,

assessed by phosphorylation of the HGF receptor (c-Met) and extracellular

signal-regulated kinase, and scattered morphology in human hepatocellular

carcinoma cell line HepG2. These results suggest that TMPRSS13

functions as an HGF-converting protease, the activity of which may be

regulated by HAI-1.

Abbreviations

BSA, bovine serum albumin; ERK, extracellular signal-regulated kinase; HA, haemagglutinin; HAI-1, hepatocyte growth factor activator

inhibitor type 1; HAI-2, hepatocyte growth factor activator inhibitor type 2; HGF, hepatocyte growth factor; HGFA, hepatocyte growth factor

activator; HPAI, highly pathogenic avian influenza; IC50, the concentration of inhibitor that inhibited the enzymatic activity by 50% compared

with the uninhibited control; LDL, low-density lipoprotein; MSPL, mosaic serine protease large form; PBS, phosphate-buffered saline;

TTSP, type II transmembrane serine protease.

4888 FEBS Journal 277 (2010) 4888–4900 ª 2010 The Authors Journal compilation ª 2010 FEBS