Mini-alphaB-crystallin: a functional element of alphaB-crystallin with chaperone-like activity.
Alpha-crystallin is a member of the family of small heat-shock proteins (sHSP) and is composed of two subunits, alphaA-crystallin and alphaB-crystallin, which exhibit molecular chaperone-like properties. In a previous study, we found that residues 70-88 in alphaA-crystallin can function like a molecular chaperone by preventing the aggregation and precipitation of denaturing substrate proteins [Sharma, K. K., et al. (2000) J. Biol. Chem. 275, 3767-3771]. In this study, we show that the complementary sequence in alphaB-crystallin, residues 73-92 (DRFSVNLDVKHFSPEELKVK), is the functional chaperone site of alphaB-crystallin. Like the mini-alphaA-crystallin chaperone, the mini-alphaB-crystallin chaperone interacts with 1,1'-bi(4-anilino) naphthalene-5,5'-disulphonic acid (bis-ANS) and also possesses significant beta-sheet and random coil structure. Deletion of four residues (DRFS) from the N-terminus or deletion of C-terminus LKVK residues from the 73-92 peptide abolishes the chaperone-like activity against denaturing alcohol dehydrogenase. However, removal of DRFS or HFSPEELKVK is necessary to completely abolish the antiaggregation property of the peptide in insulin reduction assay. Substitution of Asp at a site corresponding to D80 in alphaB-crystallin with d-Asp or beta-Asp results in a significant loss of chaperone-like activity. Kynurenine modification of His in the peptide abolishes the antiaggregation property of the mini-chaperone. These data suggest that the 73-92 region in alphaB-crystallin is one of the substrate binding sites during chaperone activity.
Bhattacharyya
Jaya
J
Department of Ophthalmology, University of Missouri, Columbia, Missouri 65212, USA.
Padmanabha Udupa
E G
EG
Wang
Jing
J
Sharma
K Krishna
KK
D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000378:metabolism; D026603:D-Aspartic Acid / Q000378:metabolism; D006639:Histidine / Q000378:metabolism; D007737:Kynurenine / Q000378:metabolism; D018832:Molecular Chaperones / Q000502:physiology; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D011489:Protein Denaturation; D013997:Time Factors; D038203:alpha-Crystallin B Chain / Q000737:chemistry / Q000502:physiology*
D018832:Molecular Chaperones; D010455:Peptides; D038203:alpha-Crystallin B Chain; D001224:Aspartic Acid; D007737:Kynurenine; D006639:Histidine; D026603:D-Aspartic Acid