Feeding recombinant E. coli with GST-mBmKTX fusion protein increases the fecundity and lifespan of Caenorhabditis elegans

Peptides. 2017 Mar:89:1-8. doi: 10.1016/j.peptides.2017.01.003. Epub 2017 Jan 11.

Abstract

Scorpion venom could be a useful treatment for a variety of diseases, such as cancer, epilepsy and analgesia. BmKTX is a polypeptide extracts from scorpion venom (PESV), which have attracted much attention from researchers in recent years. mBmKTX is a mutant polypeptide according to the amino acid sequence of BmKTX. We expressed it with the vector pGEX-4T-1 in Escherichia coli, and Caenorhabditis elegans were used as the animal model and fed with the strains. In this study, the expression of pGEX-mBmKTX was analyzed by SDS-PAGE, and GST-mBmKTX purified from pGEX-mBmKTX as a glutathione S-transferase (GST)-tagged fusion protein is approximately 30kDa. The secondary structure prediction shows that mBmKTX is mainly composed of approximately 13% β-sheet and 86% loop. A food clearance assay and brood size assay indicated that the worms fed pGEX-mBmKTX ate more and had greater fecundity than those fed the empty vector. A lifespan analysis demonstrated that mBmKTX could significantly prolong the lifespan of C. elegans, with an increase of 22.5% compared with the control. Behavioral assays confirmed that mBmKTX had no influence on the locomotion of C. elegans. In addition, microarray analysis and quantitative real-time PCR demonstrated that there are 320 differentially expressed genes, 182 of which are related to reproduction, growth and lifespan. In conclusion, the data suggested that mBmKTX has potential utility for increasing fecundity and animal survival.

Keywords: C. elegans; Fecundity; Lifespan; Microarray; mBmKTX.

MeSH terms

  • Amino Acid Sequence / genetics
  • Animals
  • Caenorhabditis elegans / drug effects
  • Caenorhabditis elegans / genetics
  • Caenorhabditis elegans Proteins / biosynthesis
  • Escherichia coli / genetics
  • Fertility / drug effects
  • Fertility / genetics
  • Gene Expression / drug effects
  • Glutathione Transferase / genetics*
  • Glutathione Transferase / pharmacology
  • Longevity / drug effects
  • Longevity / genetics
  • Microarray Analysis
  • Mutant Proteins / genetics*
  • Mutant Proteins / pharmacology
  • Peptides / genetics*
  • Peptides / pharmacology
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / pharmacology
  • Scorpion Venoms / genetics*
  • Scorpion Venoms / pharmacology

Substances

  • Caenorhabditis elegans Proteins
  • KTX toxin, Buthus
  • Mutant Proteins
  • Peptides
  • Recombinant Fusion Proteins
  • Scorpion Venoms
  • Glutathione Transferase