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Identification of LMAN1 as a Druggable Cellular Target for Reduced Secretion of Matrix Metalloproteinase‐9
Author(s) -
Duellman Tyler,
Burnett Jack,
Yang Jay
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.1189.6
Subject(s) - secretion , golgi apparatus , endoplasmic reticulum , secretory pathway , microbiology and biotechnology , secretory protein , protein fragment complementation assay , biology , matrix metalloproteinase , glycoprotein , complementation , biochemistry , gene , phenotype
Matrix metalloproteinase‐9 (MMP‐9) is a secreted endopeptidase playing a major role in shaping the extracellular environment. Elevated levels of secreted MMP‐9 have been implicated in the pathogenesis of a number of diseases and thus a detailed understanding of the secretory mechanism of this enzyme could help identify novel cellular targets to prevent or reverse disease conditions that result from excess levels of secreted MMP‐9. Previous reports have characterized some of the early secretory events of MMP‐9 showing that this glycoprotein follows the canonical secretory pathway entering the quality control folding cycle in the endoplasmic reticulum (ER) before transport of the properly folded protein to the Golgi apparatus and beyond for secretion. Despite this knowledge there is limited information about the other secretory events including the mechanism by which MMP‐9 exits the ER. CRISPR‐mediated knockout of the ER resident carrier protein LMAN1 (Lectin, Mannose‐Binding 1) confirm its functional role in the efficient secretion of MMP‐9. Through a complementation assay we determined that LMAN1, a well‐studied lectin‐carrier protein, interacts with the secretion‐competent fully N‐glycosylated MMP‐9 in the ER. Our study suggests LMAN1 selectively interacts with the properly folded, fully N‐glycosylated MMP‐9 within the ER and shuttles it to the ER‐Golgi Intermediate Compartment (ERGIC). This study has identified a novel cellular target in LMAN1 that can be therapeutically targeted for inhibition of MMP‐9 secretion. Importantly the MMP‐9: LMAN1 complementation assay provides a valuable tool for the development of a fluorescence‐based high‐throughput drug screening assay aimed at finding small molecules capable of interfering with the LMAN1: MMP‐9 interaction and thus reducing the secretion of MMP‐9. Support or Funding Information This study was supported by the Clinical Translational Cardiovascular Training Grant T32 HL07936‐12 (TD), Bamforth Endowment Fund (JY) from the Department of Anesthesiology, UW Madison, and NIH RO1 GM107054 and GM105665 (JY).