Global structural changes in annexin 12. The roles of phospholipid, Ca2+, and pH.

Isas JM, Patel DR, Jao C, Jayasinghe S, Cartailler JP, Haigler HT, Langen R
J Biol Chem. 2003 278 (32): 30227-34

PMID: 12756261 · DOI:10.1074/jbc.M301228200

Ca2+-dependent membrane interaction has long been recognized as a general property of the annexin (ANX) family of proteins. More recently, it has become clear that ANXs can also undergo Ca2+-independent membrane interactions at mildly acidic pH. Here we use site-directed spin labeling in combination with circular dichroism and biochemical labeling methods to compare the structure and membrane topography of these two different membrane-bound forms of ANX12. Our results reveal strong similarities between the solution structure and the structure of the Ca2+-dependent membrane-bound form at neutral pH. In contrast, all Ca2+-independent membrane interactions tested resulted in large scale conformational changes and membrane insertion. Pairs of spin labels that were in close proximity across the interface of different domains of the protein in both the soluble and Ca2+-dependent membrane form were >25 A apart in the Ca2+-independent membrane-bound form. Despite these major conformational changes, the overall secondary structure content did not appear to be strongly altered and ANX12 remained largely helical. Thus, Ca2+-independent membrane interaction leads to massive refolding but not unfolding. Refolding did not occur at low pH in the absence of membranes but occurred within a few seconds after phospholipid vesicles were added. The phospholipid composition of the vesicles was an important modulator of Ca2+-independent membrane interaction. For example, cardiolipin-containing vesicles induced Ca2+-independent membrane interaction even at near neutral pH, thereby raising the possibility that lipid composition could induce relatively rapid Ca2+-independent membrane interaction in vivo.

MeSH Terms (21)

Animals Annexins Calcium Cardiolipins Cattle Cell Membrane Circular Dichroism Crystallography, X-Ray Electron Spin Resonance Spectroscopy Hydrogen-Ion Concentration Kinetics Models, Molecular Mutation Phospholipids Protein Conformation Protein Denaturation Protein Folding Protein Structure, Secondary Spin Labels Time Factors Ultraviolet Rays

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