What can be learned about the function of a single protein from its various X-ray structures: the example of the sarcoplasmic calcium pump

Methods Mol Biol. 2010:654:119-40. doi: 10.1007/978-1-60761-762-4_7.

Abstract

Improvements in the handling of membrane proteins for crystallization, combined with better synchrotron sources for X-ray diffraction analysis, are leading to clarification of the structural details of an ever increasing number of membrane transporters and receptors. Here we describe how this development has resulted in the elucidation at atomic resolution of a large number of structures of the sarcoplasmic Ca(2+)-ATPase (SERCA1a) present in skeletal muscle. The structures corresponding to the various intermediary states have been obtained after stabilization with structural analogues of ATP and of metal fluorides as mimicks of inorganic phosphate. From these results it is possible, in accordance with previous biochemical and molecular biology data, to give a detailed structural description of both ATP hydrolysis and Ca(2+) transport through the membrane, to serve as the starting point for a fuller understanding of the pump mechanism and, in future studies, on the regulatory role of this ubiquitous intracellular Ca(2+)-ATPase in cellular Ca(2+) metabolism in normal and pathological conditions.

MeSH terms

  • Biological Transport
  • Calcium / metabolism
  • Calcium-Transporting ATPases / chemistry*
  • Calcium-Transporting ATPases / metabolism*
  • Fluorides / metabolism
  • Models, Biological
  • Protein Structure, Secondary
  • Sarcoplasmic Reticulum / metabolism*
  • X-Ray Diffraction / methods*

Substances

  • Calcium-Transporting ATPases
  • Fluorides
  • Calcium