LMAN1 (ERGIC-53) is a potential carrier protein for matrix metalloproteinase-9 glycoprotein secretion

Biochem Biophys Res Commun. 2015 Aug 28;464(3):685-91. doi: 10.1016/j.bbrc.2015.06.164. Epub 2015 Jul 3.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Chloroquine / pharmacology
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Endoplasmic Reticulum / metabolism
  • Gene Knockout Techniques
  • Genetic Complementation Test
  • Glycosylation
  • HEK293 Cells
  • Humans
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mannose-Binding Lectins / deficiency
  • Mannose-Binding Lectins / genetics
  • Mannose-Binding Lectins / metabolism*
  • Matrix Metalloproteinase 9 / chemistry
  • Matrix Metalloproteinase 9 / metabolism*
  • Membrane Proteins / deficiency
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mutagenesis, Site-Directed
  • Protein Interaction Domains and Motifs
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Bacterial Proteins
  • LMAN1 protein, human
  • Luminescent Proteins
  • Mannose-Binding Lectins
  • Membrane Proteins
  • Recombinant Fusion Proteins
  • yellow fluorescent protein, Bacteria
  • Chloroquine
  • MMP9 protein, human
  • Matrix Metalloproteinase 9