Abstract
Matrix metalloproteinase-9 (MMP-9) is a secreted glycoprotein with a major role in shaping the extracellular matrix and a detailed understanding of the secretory mechanism could help identify methods to correct diseases resulting from dysregulation of secretion. MMP-9 appears to follow a canonical secretory pathway through a quality control cycle in the endoplasmic reticulum (ER) before transport of the properly folded protein to the Golgi apparatus and beyond for secretion. Through a complementation assay, we determined that LMAN1, a well-studied lectin-carrier protein, interacts with a secretion-competent N-glycosylated MMP-9 in the ER while N-glycosylation-deficient secretion-compromised MMP-9 does not. In contrast, co-immunoprecipitation demonstrated protein interaction between LMAN1 and secretion-compromised N-glycosylation-deficient MMP-9. MMP-9 secretion was reduced in the LMAN1 knockout cell line compared to control cells confirming the functional role of LMAN1. These observations support the role of LMAN1 as a lectin-carrier protein mediating efficient MMP-9 secretion.
Keywords:
Glycoprotein; Lectin-carrier protein; Matrix metalloproteinase-9; Secretion.
Copyright © 2015 Elsevier Inc. All rights reserved.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Substitution
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Bacterial Proteins / chemistry
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism
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Chloroquine / pharmacology
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Clustered Regularly Interspaced Short Palindromic Repeats
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Endoplasmic Reticulum / metabolism
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Gene Knockout Techniques
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Genetic Complementation Test
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Glycosylation
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HEK293 Cells
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Humans
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Luminescent Proteins / chemistry
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Mannose-Binding Lectins / deficiency
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Mannose-Binding Lectins / genetics
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Mannose-Binding Lectins / metabolism*
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Matrix Metalloproteinase 9 / chemistry
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Matrix Metalloproteinase 9 / metabolism*
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Membrane Proteins / deficiency
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Mutagenesis, Site-Directed
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Protein Interaction Domains and Motifs
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
Substances
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Bacterial Proteins
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LMAN1 protein, human
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Luminescent Proteins
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Mannose-Binding Lectins
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Membrane Proteins
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Recombinant Fusion Proteins
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yellow fluorescent protein, Bacteria
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Chloroquine
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MMP9 protein, human
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Matrix Metalloproteinase 9