In Vitro Comparison of Adipokine Export Signals

Title:In Vitro Comparison of Adipokine Export Signals

Volume: 23 Issue: 6

Author(s): Parisa Sharafi and Y. Çetin Kocaefe

Affiliation:

Keywords: Adipocytes, adipokines, export signals, recombinant protein export.

Abstract: Mammalian cells are widely used for recombinant protein production in research and biotechnology. Utilization of export signals significantly facilitates production and purification processes. 35 years after the discovery of the mammalian export machinery, there still are obscurities regarding the efficiency of the export signals. The aim of this study was the comparative evaluation of the efficiency of selected export signals using adipocytes as a cell model. Adipocytes have a large capacity for protein secretion including several enzymes, adipokines, and other signaling molecules, providing a valid system for a quantitative evaluation. Constructs that expressed N-terminal fusion export signals were generated to express Enhanced Green Fluorescence Protein (EGFP) as a reporter for quantitative and qualitative evaluation. Furthermore, fluorescent microscopy was used to trace the intracellular traffic of the reporter. The export efficiency of six selected proteins secreted from adipocytes was evaluated. Quantitative comparison of intracellular and exported fractions of the recombinant constructs demonstrated a similar efficiency among the studied sequences with minor variations. The export signal of Retinol Binding Protein (RBP4) exhibited the highest efficiency. This study presents the first quantitative data showing variations among export signals, in adipocytes which will help optimization of recombinant protein distribution.

Cite this article as:

Sharafi Parisa and Çetin Kocaefe Y., In Vitro Comparison of Adipokine Export Signals, Protein & Peptide Letters 2016; 23 (6) . https://dx.doi.org/10.2174/0929866523666160411144531