Interactive domains for chaperone activity in the small heat shock protein, human alphaB crystallin.

Biochemistry

Biomolecular Structure and Design, University of Washington, Seattle, Washington 98195-7420, USA.

Published: November 2005

AI Article Synopsis

  • Protein pin arrays discovered seven sequences that indicate chaperone activity in human alphaB crystallin, utilizing natural lens proteins and in vitro targets.
  • The N-terminal domain had two interactive sequences, while the alpha crystallin core domain featured four sequences, and one was found in the C-terminal domain.
  • Synthesized peptides from the identified sequences successfully inhibited thermal aggregation of key proteins, demonstrating their role in chaperone activity and suggesting these sequences also contribute to subunit assembly in alphaB crystallin.

Article Abstract

Protein pin arrays identified seven interactive sequences for chaperone activity in human alphaB crystallin using natural lens proteins, beta(H) crystallin and gammaD crystallin, and in vitro chaperone target proteins, alcohol dehydrogenase and citrate synthase. The N-terminal domain contained two interactive sequences, (9)WIRRPFFPFHSP(20) and (43)SLSPFYLRPPSFLRAP(58). The alpha crystallin core domain contained four interactive sequences, (75)FSVNLDVK(82) (beta3), (113)FISREFHR(120), (131)LTITSSLS(138) (beta8), and (141)GVLTVNGP(148) (beta9). The C-terminal domain contained one interactive sequence, (157)RTIPITRE(164), that included the highly conserved I-X-I/V motif. Two interactive sequences, (73)DRFSVNLDVKHFS(85) and (131)LTITSSLSDGV(141), belonging to the alpha crystallin core domain were synthesized as peptides and assayed for chaperone activity in vitro. Both synthesized peptides inhibited the thermal aggregation of beta(H) crystallin, alcohol dehydrogenase, and citrate synthase in vitro. Five of the seven chaperone sequences identified by the pin arrays overlapped with sequences identified previously as sequences for subunit-subunit interactions in human alphaB crystallin. The results suggested that interactive sequences in human alphaB crystallin have dual roles in subunit-subunit assembly and chaperone activity.

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http://dx.doi.org/10.1021/bi0503910DOI Listing

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