Unconventional secretion of the mutated 19 kDa protein of Oplophorus luciferase (nanoKAZ) in mammalian cells

Biochem Biophys Res Commun. 2014 Aug 8;450(4):1313-9. doi: 10.1016/j.bbrc.2014.06.140. Epub 2014 Jul 11.

Abstract

The putative amino-terminal signal peptide of the catalytic 19 kDa protein (KAZ) of Oplophorus luciferase was found to be a functional secretory peptide in mammalian cells. A 16 amino acid substituted mutant of KAZ (nanoKAZ) could be secreted from mammalian cells using the amino-terminal signal peptide of KAZ, but KAZ could not be secreted at all. Notably, nanoKAZ lacking the amino-terminal signal peptide could be secreted from mammalian cells, and the distribution of nanoKAZ on the cell membrane was confirmed by video-rate bioluminescence imaging. Thus, nanoKAZ lacking the amino-terminal signal peptide was expressed in the cytoplasm, translocated to the cell membrane, and released into the culture medium through an endoplasmic reticulum-Golgi-independent pathway.

Keywords: Amino-terminal signal peptide; Bioluminescence imaging; Brefeldin A; ER/Golgi-dependent secretion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arthropod Proteins / chemistry
  • Arthropod Proteins / genetics
  • Arthropod Proteins / metabolism*
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Culture Media
  • Decapoda / enzymology*
  • Electrophoresis, Polyacrylamide Gel
  • Luciferases / metabolism*
  • Molecular Sequence Data
  • Mutation*
  • Protein Sorting Signals
  • Sequence Homology, Amino Acid

Substances

  • Arthropod Proteins
  • Culture Media
  • Protein Sorting Signals
  • Luciferases