The N-terminal ectodomain of Ninjurin1 liberated by MMP9 has chemotactic activity

Bum Ju Ahn; Hoang Le; Min Wook Shin; Sung Jin Bae; Eun Ji Lee; Hee Jun Wee; Jong Ho Cha; Ji Hyeon Park; Hye Shin Lee; Hyo Jong Lee; Hyunsook Jung; Zee Yong Park; Sang Ho Park; Byung Woo Han; Ji Hae Seo; Eng H. Lo; Kyu Won Kim

doi:10.1016/j.bbrc.2012.10.099

The N-terminal ectodomain of Ninjurin1 liberated by MMP9 has chemotactic activity

Bum Ju Ahn, Hoang Le, Min Wook Shin, Sung Jin Bae, Eun Ji Lee, Hee Jun Wee, Jong Ho Cha, Ji Hyeon Park, Hye Shin Lee, Hyo Jong Lee, Hyunsook Jung, Zee Yong Park, Sang Ho Park, Byung Woo Han, Ji Hae Seo, Eng H. Lo, Kyu Won Kim

Research output: Contribution to journal › Article › peer-review

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Abstract

Ninjurin1 is known as an adhesion molecule promoting leukocyte trafficking under inflammatory conditions. However, the posttranslational modifications of Ninjurin1 are poorly understood. Herein, we defined the proteolytic cleavage of Ninjurin1 and its functions. HEK293T cells overexpressing the C- or N-terminus tagging mouse Ninjurin1 plasmid produced additional cleaved forms of Ninjurin1 in the lysates or conditioned media (CM). Two custom-made anti-Ninjurin1 antibodies, Ab_1-15 or Ab_139-152, specific to the N- or C-terminal regions of Ninjurin1 revealed the presence of its shedding fragments in the mouse liver and kidney lysates. Furthermore, Matrix Metalloproteinase (MMP) 9 was responsible for Ninjurin1 cleavage between Leu⁵⁶ and Leu⁵⁷. Interestingly, the soluble N-terminal Ninjurin1 fragment has structural similarity with well-known chemokines. Indeed, the CM from HEK293T cells overexpressing the GFP-mNinj1 plasmid was able to attract Raw264.7 cells in trans-well assay. Collectively, we suggest that the N-terminal ectodomain of mouse Ninjurin1, which may act as a chemoattractant, is cleaved by MMP9.

Original language	English
Pages (from-to)	438-444
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	428
Issue number	4
DOIs	https://doi.org/10.1016/j.bbrc.2012.10.099
State	Published - 30 Nov 2012
Externally published	Yes

Bibliographical note

Funding Information:
This work was supported by a grant from the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (MEST) through the Creative Research Initiative Program ( R16-2004-001-01001-0 ), the World Class University Program ( R31-2008-000-10103-0 ), the Global Research Laboratory Program ( 2011-0021874 ), and the Global Core Research Center (GCRC) Program ( 2012-0001187 ). Z.-Y Park was supported by the “2012 GIST Basic Research Projects in High-tech Industrial Technology”. E.H. Lo was supported by grants from National Institutes of Health (R37-NS37074, R01-76694 and P01-NS55104).

Keywords

Chemotaxis
MMP9
Ninjurin1
Posttranslational modification
Proteolytic cleavage

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10.1016/j.bbrc.2012.10.099

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Ahn, B. J., Le, H., Shin, M. W., Bae, S. J., Lee, E. J., Wee, H. J., Cha, J. H., Park, J. H., Lee, H. S., Lee, H. J., Jung, H., Park, Z. Y., Park, S. H., Han, B. W., Seo, J. H., Lo, E. H., & Kim, K. W. (2012). The N-terminal ectodomain of Ninjurin1 liberated by MMP9 has chemotactic activity. Biochemical and Biophysical Research Communications, 428(4), 438-444. https://doi.org/10.1016/j.bbrc.2012.10.099

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title = "The N-terminal ectodomain of Ninjurin1 liberated by MMP9 has chemotactic activity",

abstract = "Ninjurin1 is known as an adhesion molecule promoting leukocyte trafficking under inflammatory conditions. However, the posttranslational modifications of Ninjurin1 are poorly understood. Herein, we defined the proteolytic cleavage of Ninjurin1 and its functions. HEK293T cells overexpressing the C- or N-terminus tagging mouse Ninjurin1 plasmid produced additional cleaved forms of Ninjurin1 in the lysates or conditioned media (CM). Two custom-made anti-Ninjurin1 antibodies, Ab1-15 or Ab139-152, specific to the N- or C-terminal regions of Ninjurin1 revealed the presence of its shedding fragments in the mouse liver and kidney lysates. Furthermore, Matrix Metalloproteinase (MMP) 9 was responsible for Ninjurin1 cleavage between Leu56 and Leu57. Interestingly, the soluble N-terminal Ninjurin1 fragment has structural similarity with well-known chemokines. Indeed, the CM from HEK293T cells overexpressing the GFP-mNinj1 plasmid was able to attract Raw264.7 cells in trans-well assay. Collectively, we suggest that the N-terminal ectodomain of mouse Ninjurin1, which may act as a chemoattractant, is cleaved by MMP9.",

keywords = "Chemotaxis, MMP9, Ninjurin1, Posttranslational modification, Proteolytic cleavage",

author = "Ahn, {Bum Ju} and Hoang Le and Shin, {Min Wook} and Bae, {Sung Jin} and Lee, {Eun Ji} and Wee, {Hee Jun} and Cha, {Jong Ho} and Park, {Ji Hyeon} and Lee, {Hye Shin} and Lee, {Hyo Jong} and Hyunsook Jung and Park, {Zee Yong} and Park, {Sang Ho} and Han, {Byung Woo} and Seo, {Ji Hae} and Lo, {Eng H.} and Kim, {Kyu Won}",

note = "Funding Information: This work was supported by a grant from the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (MEST) through the Creative Research Initiative Program ( R16-2004-001-01001-0 ), the World Class University Program ( R31-2008-000-10103-0 ), the Global Research Laboratory Program ( 2011-0021874 ), and the Global Core Research Center (GCRC) Program ( 2012-0001187 ). Z.-Y Park was supported by the “2012 GIST Basic Research Projects in High-tech Industrial Technology”. E.H. Lo was supported by grants from National Institutes of Health (R37-NS37074, R01-76694 and P01-NS55104).",

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Ahn, BJ, Le, H, Shin, MW, Bae, SJ, Lee, EJ, Wee, HJ, Cha, JH, Park, JH, Lee, HS, Lee, HJ, Jung, H, Park, ZY, Park, SH, Han, BW, Seo, JH, Lo, EH & Kim, KW 2012, 'The N-terminal ectodomain of Ninjurin1 liberated by MMP9 has chemotactic activity', Biochemical and Biophysical Research Communications, vol. 428, no. 4, pp. 438-444. https://doi.org/10.1016/j.bbrc.2012.10.099

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T1 - The N-terminal ectodomain of Ninjurin1 liberated by MMP9 has chemotactic activity

AU - Ahn, Bum Ju

AU - Le, Hoang

AU - Shin, Min Wook

AU - Bae, Sung Jin

AU - Lee, Eun Ji

AU - Wee, Hee Jun

AU - Cha, Jong Ho

AU - Park, Ji Hyeon

AU - Lee, Hye Shin

AU - Lee, Hyo Jong

AU - Jung, Hyunsook

AU - Park, Zee Yong

AU - Park, Sang Ho

AU - Han, Byung Woo

AU - Seo, Ji Hae

AU - Lo, Eng H.

AU - Kim, Kyu Won

N1 - Funding Information: This work was supported by a grant from the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (MEST) through the Creative Research Initiative Program ( R16-2004-001-01001-0 ), the World Class University Program ( R31-2008-000-10103-0 ), the Global Research Laboratory Program ( 2011-0021874 ), and the Global Core Research Center (GCRC) Program ( 2012-0001187 ). Z.-Y Park was supported by the “2012 GIST Basic Research Projects in High-tech Industrial Technology”. E.H. Lo was supported by grants from National Institutes of Health (R37-NS37074, R01-76694 and P01-NS55104).

PY - 2012/11/30

Y1 - 2012/11/30

N2 - Ninjurin1 is known as an adhesion molecule promoting leukocyte trafficking under inflammatory conditions. However, the posttranslational modifications of Ninjurin1 are poorly understood. Herein, we defined the proteolytic cleavage of Ninjurin1 and its functions. HEK293T cells overexpressing the C- or N-terminus tagging mouse Ninjurin1 plasmid produced additional cleaved forms of Ninjurin1 in the lysates or conditioned media (CM). Two custom-made anti-Ninjurin1 antibodies, Ab1-15 or Ab139-152, specific to the N- or C-terminal regions of Ninjurin1 revealed the presence of its shedding fragments in the mouse liver and kidney lysates. Furthermore, Matrix Metalloproteinase (MMP) 9 was responsible for Ninjurin1 cleavage between Leu56 and Leu57. Interestingly, the soluble N-terminal Ninjurin1 fragment has structural similarity with well-known chemokines. Indeed, the CM from HEK293T cells overexpressing the GFP-mNinj1 plasmid was able to attract Raw264.7 cells in trans-well assay. Collectively, we suggest that the N-terminal ectodomain of mouse Ninjurin1, which may act as a chemoattractant, is cleaved by MMP9.

AB - Ninjurin1 is known as an adhesion molecule promoting leukocyte trafficking under inflammatory conditions. However, the posttranslational modifications of Ninjurin1 are poorly understood. Herein, we defined the proteolytic cleavage of Ninjurin1 and its functions. HEK293T cells overexpressing the C- or N-terminus tagging mouse Ninjurin1 plasmid produced additional cleaved forms of Ninjurin1 in the lysates or conditioned media (CM). Two custom-made anti-Ninjurin1 antibodies, Ab1-15 or Ab139-152, specific to the N- or C-terminal regions of Ninjurin1 revealed the presence of its shedding fragments in the mouse liver and kidney lysates. Furthermore, Matrix Metalloproteinase (MMP) 9 was responsible for Ninjurin1 cleavage between Leu56 and Leu57. Interestingly, the soluble N-terminal Ninjurin1 fragment has structural similarity with well-known chemokines. Indeed, the CM from HEK293T cells overexpressing the GFP-mNinj1 plasmid was able to attract Raw264.7 cells in trans-well assay. Collectively, we suggest that the N-terminal ectodomain of mouse Ninjurin1, which may act as a chemoattractant, is cleaved by MMP9.

KW - Chemotaxis

KW - MMP9

KW - Ninjurin1

KW - Posttranslational modification

KW - Proteolytic cleavage

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SN - 0006-291X

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JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

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The N-terminal ectodomain of Ninjurin1 liberated by MMP9 has chemotactic activity

Abstract

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Keywords

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