Hepatocyte growth factor activator inhibitor-2 prevents shedding of matriptase

Brian R Larsen; Simon D R Steffensen; Nis Valentin Ladefoged Nielsen; Stine Friis; Sine Godiksen; Jette Bornholdt Lange; Christoffer Soendergaard; Annika W Nonboe; Martin N Andersen; Steen S Poulsen; Roman Szabo; Thomas H Bugge; Chen-Yong Lin; Hanne Skovbjerg; Jan K Jensen; Lotte K Vogel

doi:10.1016/j.yexcr.2013.01.008

Hepatocyte growth factor activator inhibitor-2 prevents shedding of matriptase

Brian R Larsen, Simon D R Steffensen, Nis Valentin Ladefoged Nielsen, Stine Friis, Sine Godiksen, Jette Bornholdt Lange, Christoffer Soendergaard, Annika W Nonboe, Martin N Andersen, Steen S Poulsen, Roman Szabo, Thomas H Bugge, Chen-Yong Lin, Hanne Skovbjerg, Jan K Jensen, Lotte K Vogel

26 Citations (Scopus)

Abstract

Hepatocyte growth factor activator inhibitor-2 (HAI-2) is an inhibitor of many proteases in vitro, including the membrane-bound serine protease, matriptase. Studies of knock-out mice have shown that HAI-2 is essential for placental development only in mice expressing matriptase, suggesting that HAI-2 is important for regulation of matriptase. Previous studies have shown that recombinant expression of matriptase was unsuccessful unless co-expressed with another HAI, HAI-1. In the present study we show that when human matriptase is recombinantly expressed alone in the canine cell line MDCK, then human matriptase mRNA can be detected and the human matriptase ectodomain is shed to the media, suggesting that matriptase expressed alone is rapidly transported through the secretory pathway and shed. Whereas matriptase expressed together with HAI-1 or HAI-2 accumulates on the plasma membrane where it is activated, as judged by cleavage at Arg614 and increased peptidolytic activity of the cell extracts. Mutagenesis of Kunitz domain 1 but not Kunitz domain 2 abolished this function of HAI-2. HAI-2 seems to carry out its function intracellularly as this is where the vast majority of HAI-2 is located and since HAI-2 could not be detected on the basolateral plasma membrane where matriptase resides. However, minor amounts of HAI-2 not undergoing endocytosis could be detected on the apical plasma membrane. Our results suggest that Kunitz domain 1 of HAI-2 cause matriptase to accumulate in a membrane-bound form on the basolateral plasma membrane.

Original language	English
Journal	Experimental Cell Research
Volume	319
Issue number	6
Pages (from-to)	918-929
Number of pages	12
ISSN	0014-4827
DOIs	https://doi.org/10.1016/j.yexcr.2013.01.008
Publication status	Published - 1 Apr 2013

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Larsen, B. R., Steffensen, S. D. R., Nielsen, N. V. L., Friis, S., Godiksen, S., Lange, J. B., Soendergaard, C., Nonboe, A. W., Andersen, M. N., Poulsen, S. S., Szabo, R., Bugge, T. H., Lin, C-Y., Skovbjerg, H., Jensen, J. K., & Vogel, L. K. (2013). Hepatocyte growth factor activator inhibitor-2 prevents shedding of matriptase. Experimental Cell Research, 319(6), 918-929. https://doi.org/10.1016/j.yexcr.2013.01.008

Larsen, BR, Steffensen, SDR, Nielsen, NVL, Friis, S, Godiksen, S, Lange, JB, Soendergaard, C, Nonboe, AW, Andersen, MN, Poulsen, SS, Szabo, R, Bugge, TH, Lin, C-Y, Skovbjerg, H, Jensen, JK & Vogel, LK 2013, 'Hepatocyte growth factor activator inhibitor-2 prevents shedding of matriptase', Experimental Cell Research, vol. 319, no. 6, pp. 918-929. https://doi.org/10.1016/j.yexcr.2013.01.008

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title = "Hepatocyte growth factor activator inhibitor-2 prevents shedding of matriptase",

abstract = "Hepatocyte growth factor activator inhibitor-2 (HAI-2) is an inhibitor of many proteases in vitro, including the membrane-bound serine protease, matriptase. Studies of knock-out mice have shown that HAI-2 is essential for placental development only in mice expressing matriptase, suggesting that HAI-2 is important for regulation of matriptase. Previous studies have shown that recombinant expression of matriptase was unsuccessful unless co-expressed with another HAI, HAI-1. In the present study we show that when human matriptase is recombinantly expressed alone in the canine cell line MDCK, then human matriptase mRNA can be detected and the human matriptase ectodomain is shed to the media, suggesting that matriptase expressed alone is rapidly transported through the secretory pathway and shed. Whereas matriptase expressed together with HAI-1 or HAI-2 accumulates on the plasma membrane where it is activated, as judged by cleavage at Arg614 and increased peptidolytic activity of the cell extracts. Mutagenesis of Kunitz domain 1 but not Kunitz domain 2 abolished this function of HAI-2. HAI-2 seems to carry out its function intracellularly as this is where the vast majority of HAI-2 is located and since HAI-2 could not be detected on the basolateral plasma membrane where matriptase resides. However, minor amounts of HAI-2 not undergoing endocytosis could be detected on the apical plasma membrane. Our results suggest that Kunitz domain 1 of HAI-2 cause matriptase to accumulate in a membrane-bound form on the basolateral plasma membrane.",

author = "Larsen, {Brian R} and Steffensen, {Simon D R} and Nielsen, {Nis Valentin Ladefoged} and Stine Friis and Sine Godiksen and Lange, {Jette Bornholdt} and Christoffer Soendergaard and Nonboe, {Annika W} and Andersen, {Martin N} and Poulsen, {Steen S} and Roman Szabo and Bugge, {Thomas H} and Chen-Yong Lin and Hanne Skovbjerg and Jensen, {Jan K} and Vogel, {Lotte K}",

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doi = "10.1016/j.yexcr.2013.01.008",

language = "English",

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T1 - Hepatocyte growth factor activator inhibitor-2 prevents shedding of matriptase

AU - Larsen, Brian R

AU - Steffensen, Simon D R

AU - Nielsen, Nis Valentin Ladefoged

AU - Friis, Stine

AU - Godiksen, Sine

AU - Lange, Jette Bornholdt

AU - Soendergaard, Christoffer

AU - Nonboe, Annika W

AU - Andersen, Martin N

AU - Poulsen, Steen S

AU - Szabo, Roman

AU - Bugge, Thomas H

AU - Lin, Chen-Yong

AU - Skovbjerg, Hanne

AU - Jensen, Jan K

AU - Vogel, Lotte K

PY - 2013/4/1

Y1 - 2013/4/1

N2 - Hepatocyte growth factor activator inhibitor-2 (HAI-2) is an inhibitor of many proteases in vitro, including the membrane-bound serine protease, matriptase. Studies of knock-out mice have shown that HAI-2 is essential for placental development only in mice expressing matriptase, suggesting that HAI-2 is important for regulation of matriptase. Previous studies have shown that recombinant expression of matriptase was unsuccessful unless co-expressed with another HAI, HAI-1. In the present study we show that when human matriptase is recombinantly expressed alone in the canine cell line MDCK, then human matriptase mRNA can be detected and the human matriptase ectodomain is shed to the media, suggesting that matriptase expressed alone is rapidly transported through the secretory pathway and shed. Whereas matriptase expressed together with HAI-1 or HAI-2 accumulates on the plasma membrane where it is activated, as judged by cleavage at Arg614 and increased peptidolytic activity of the cell extracts. Mutagenesis of Kunitz domain 1 but not Kunitz domain 2 abolished this function of HAI-2. HAI-2 seems to carry out its function intracellularly as this is where the vast majority of HAI-2 is located and since HAI-2 could not be detected on the basolateral plasma membrane where matriptase resides. However, minor amounts of HAI-2 not undergoing endocytosis could be detected on the apical plasma membrane. Our results suggest that Kunitz domain 1 of HAI-2 cause matriptase to accumulate in a membrane-bound form on the basolateral plasma membrane.

AB - Hepatocyte growth factor activator inhibitor-2 (HAI-2) is an inhibitor of many proteases in vitro, including the membrane-bound serine protease, matriptase. Studies of knock-out mice have shown that HAI-2 is essential for placental development only in mice expressing matriptase, suggesting that HAI-2 is important for regulation of matriptase. Previous studies have shown that recombinant expression of matriptase was unsuccessful unless co-expressed with another HAI, HAI-1. In the present study we show that when human matriptase is recombinantly expressed alone in the canine cell line MDCK, then human matriptase mRNA can be detected and the human matriptase ectodomain is shed to the media, suggesting that matriptase expressed alone is rapidly transported through the secretory pathway and shed. Whereas matriptase expressed together with HAI-1 or HAI-2 accumulates on the plasma membrane where it is activated, as judged by cleavage at Arg614 and increased peptidolytic activity of the cell extracts. Mutagenesis of Kunitz domain 1 but not Kunitz domain 2 abolished this function of HAI-2. HAI-2 seems to carry out its function intracellularly as this is where the vast majority of HAI-2 is located and since HAI-2 could not be detected on the basolateral plasma membrane where matriptase resides. However, minor amounts of HAI-2 not undergoing endocytosis could be detected on the apical plasma membrane. Our results suggest that Kunitz domain 1 of HAI-2 cause matriptase to accumulate in a membrane-bound form on the basolateral plasma membrane.

U2 - 10.1016/j.yexcr.2013.01.008

DO - 10.1016/j.yexcr.2013.01.008

M3 - Journal article

C2 - 23333561

SN - 0014-4827

VL - 319

SP - 918

EP - 929

JO - Experimental Cell Research

JF - Experimental Cell Research

IS - 6

ER -

Hepatocyte growth factor activator inhibitor-2 prevents shedding of matriptase

Abstract

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