All You Ever Wanted to Know About APOL1 and TLFs and Did Not Dare Ask

Methods Mol Biol. 2020:2116:463-483. doi: 10.1007/978-1-0716-0294-2_28.

Abstract

Interest in trypanosome lytic factors (TLFs) and apolipoprotein L1, the ion channel-forming protein component of TLFs, has increased tenfold since 2010. This is due to the association of African variants of APOL1 with kidney disease such that interest has reached circles beyond parasitology. We have extensive experience purifying and working with these proteins and protein complexes. Herein we describe our detailed purification protocols to aid the new burgeoning field by providing an opportunity for consistency in reagents used across laboratories. We emphasize that it is imperative to maintain APOL1 protein intact (~42 kDa) to analyze the active ion channel-forming component/protein.

Keywords: Apolipoprotein L1; Haptoglobin related protein; High density lipoprotein; Ion channel; Kidney disease; Plasma; Serum; Trypanosome lytic factor.

MeSH terms

  • Apolipoprotein L1 / blood
  • Apolipoprotein L1 / chemistry
  • Apolipoprotein L1 / isolation & purification*
  • Apolipoprotein L1 / metabolism
  • Humans
  • Kidney Diseases / blood
  • Kidney Diseases / immunology
  • Lipoproteins, HDL / chemistry
  • Lipoproteins, HDL / isolation & purification*
  • Lipoproteins, HDL / metabolism
  • Molecular Weight
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Trypanosoma / immunology
  • Trypanosomiasis, African / blood*
  • Trypanosomiasis, African / complications
  • Trypanosomiasis, African / immunology
  • Trypanosomiasis, African / parasitology

Substances

  • APOL1 protein, human
  • Apolipoprotein L1
  • Lipoproteins, HDL
  • Recombinant Proteins
  • TLF1 protein, human