Crystal structures of Apo- and holo-bovine α-lactalbumin at 2.2-Å resolution reveal an effect of calcium on inter-lobe interactions

E. D. Chrysina, K. Brew, K. R. Acharya

Research output: Contribution to journalArticlepeer-review

192 Citations (SciVal)

Abstract

High affinity binding of Ca2+ to α-lactalbumin (LA) stabilizes the native structure and is required for the efficient generation of native protein with correct disulfide bonds from the reduced denatured state. A progressive increase in affinity of LA conformers for Ca2+ as they develop increasingly native structures can account for the tendency of the apo form to assume a molten globule state and the large acceleration of folding by Ca2+. To investigate the effect of calcium on structure of bovine LA, x-ray structures have been determined for crystals of the apo and holo forms at 2.2-Å resolution. In both crystal forms, which were grown at high ionic strength, the protein is in a similar global native conformation consisting of α-helical and β-subdomains separated by a cleft. Even though alternative cations and Ca2+ liganding solvent molecules are absent, removal of Ca2+ has only minor effects on the structure of the metal-binding site and a structural change was observed in the cleft on the opposite face of the molecule adjoining Tyr103 of the helical lobe and Gln54 of the β-lobe. Changes include increased separation of the lobes, loss of a buried solvent molecule near the Ca2+-binding site, and the replacement of inter- and intra-lobe H-bonds of Tyr103 by interactions with new immobilized water molecules. The more open cleft structure in the apo protein appears to be an effect of calcium binding transmitted via a change in orientation of helix H3 relative to the β-lobe to the inter-lobe interface. Calcium is well known to promote the folding of LA. The results from the comparison of apo and holo structures of LA provide high resolution structural evidence that the acceleration of folding by Ca2+ is mediated by an effect on interactions between the two subdomains.

Original languageEnglish
Pages (from-to)37021-37029
Number of pages9
JournalJournal of Biological Chemistry
Volume275
Issue number47
DOIs
Publication statusPublished - 24 Nov 2000

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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