A native Zn/Cd pumping P(1B) ATPase from natural overexpression in a hyperaccumulator plant

Biochem Biophys Res Commun. 2007 Nov 9;363(1):51-6. doi: 10.1016/j.bbrc.2007.08.105. Epub 2007 Aug 28.

Abstract

We report here the first purification of a P(1B) type ATPase, a group of transporters that occurs in bacteria, plants and animals incl. humans, from a eukaryotic organism in native state. TcHMA4 is a P(1B) type ATPase that is highly expressed in the Cd/Zn hyperaccumulator plant Thlaspi caerulescens and contains a C-terminal 9-histidine repeat. After isolation from roots, we purified TcHMA4 protein via metal affinity chromatography. The purified protein exhibited Cd- and Zn-activated ATPase activity after reconstitution into lipid vesicles, showing that it was in its native state. Gels of crude root extract and of the purified protein revealed TcHMA4-specific bands of about 50 and 60kDa, respectively, while the TcHMA4 mRNA predicts a single protein with a size of 128kDa. This indicates the occurrence of post-translational processing; the properties of the two bands were characterised by their activity and binding properties.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / chemistry*
  • Cadmium / chemistry*
  • Enzyme Activation
  • Plant Extracts / chemistry*
  • Plant Proteins / chemistry*
  • Proton Pumps / chemistry*
  • Substrate Specificity
  • Thlaspi / enzymology*
  • Up-Regulation
  • Zinc / chemistry*

Substances

  • Plant Extracts
  • Plant Proteins
  • Proton Pumps
  • Cadmium
  • Adenosine Triphosphatases
  • Zinc