Sos recruitment system for the analysis of the interaction between sulfatase-modifying factor 2 subtypes and interleukin-13

Genet Mol Res. 2013 Nov 18;12(4):5664-72. doi: 10.4238/2013.November.18.15.

Abstract

Interleukin (IL)-13 is a central mediator in allergic asthma. Our previous results have indicated that sulfatase-modifying factor 2 (SUMF2) interacts with IL-13 and inhibits its secretion. In this study, we investigated the interactions between SUMF2 subtypes and 2 types of IL-13. Wild type IL-13 (wh-IL-13) and its mutated counterpart (mh-IL-13) were analyzed and cloned into pSos yeast expression vectors. Protein was expressed in host cdc25H yeast strains. A quartet of agar growth plates was prepared for the yeast two-hybrid system, which was used to detect IL-13 and SUMF2 subtype interactions. Both yeast expression vectors, pSos/whIL-13 and pSos/whIL-13, and recombinant expression vectors for the 5 subtypes of SUMF2 (pMyr/SUMF2-Vx) were constructed. Our data showed that all of the SUMF2 subtypes bound to whIL-13 and mhIL-13 in the CytoTrap system. Five SUMF2 subtypes - SUMF2-V2, SUMF2-V3, SUMF2-V4, SUMF2-V5, and SUMF2-V7--interacted with whIL-13 and mhIL-13. These subtypes may contribute to allergic asthma by mediating IL-13 release.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Humans
  • Interleukin-13 / genetics
  • Interleukin-13 / metabolism*
  • Mutation
  • Protein Binding
  • Sulfatases / genetics
  • Sulfatases / metabolism*
  • Two-Hybrid System Techniques

Substances

  • IL13 protein, human
  • Interleukin-13
  • SUMF2 protein, human
  • Sulfatases