Alpha-crystallin regions affected by adenosine 5'-triphosphate identified by hydrogen-deuterium exchange.
ATP interaction with lens alpha-crystallins leading to enhanced chaperone activity is not yet well understood. One model for chaperone activity of small heat shock proteins proposes that ATP causes small heat shock proteins to release substrates, which are then renatured by other larger heat shock proteins. A similar role has been proposed for ATP in alpha-crystallin chaperone activity. To evaluate this model, ATP-induced structural changes of native human alpha-crystallin assemblies were determined by hydrogen-deuterium exchange. In these experiments, hydrogen-deuterium exchange, measured by mass spectrometry, gave direct evidence that ATP decreases the accessibility of amide hydrogens in multiple regions of both alphaA and alphaB. The surface encompassed by these regions is much larger than would be shielded by a single ATP, implying that multiple ATP molecules bind to each subunit and/or ATP causes a more compact alpha-crystallin structure. Such a conformational change could release a bound substrate. The regions most affected by ATP are near putative substrate binding regions of alphaA and alphaB and in the C-terminal extension of alphaB. The widespread decrease in hydrogen-deuterium exchange with particularly large decreases near substrate binding regions suggests that ATP releases substrates via both direct displacement and a global conformational change.
Hasan
Azeem
A
Department of Chemistry, University of Nebraska, Lincoln, NE 68588-0304, USA.
Smith
David L
DL
Smith
Jean B
JB
D000255:Adenosine Triphosphate / Q000737:chemistry*; D000328:Adult; D000595:Amino Acid Sequence; D003903:Deuterium / Q000737:chemistry*; D006801:Humans; D006859:Hydrogen / Q000737:chemistry*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D010449:Peptide Mapping; D038202:alpha-Crystallin A Chain / Q000032:analysis / Q000737:chemistry*; D038203:alpha-Crystallin B Chain / Q000032:analysis / Q000737:chemistry*
D010446:Peptide Fragments; D038202:alpha-Crystallin A Chain; D038203:alpha-Crystallin B Chain; D006859:Hydrogen; D000255:Adenosine Triphosphate; D003903:Deuterium