Relationship between chaperone activity and oligomeric size of recombinant human alphaA- and alphaB-crystallin: a tryptic digestion study.
Alpha-crystallin, the major eye lens protein, exists as a large oligomer of two subunits, alphaA- and alphaB-crystallin. The individual subunits assemble into the oligomer in vitro. It is generally believed that oligomerization is pre-requisite for chaperone function, although there is no hard data available on this subject. We therefore undertook a study using limited tryptic digestion as a tool for examining the relationship between oligomeric size and chaperone activity of recombinant alphaA- and alphaB-crystallin. We showed that tryptic digested fragments of both alphaA- and alphaB-crystallin much smaller than the original subunits retain considerable chaperone activity. Our results indicate that chaperone activity depends more on the sequence of the reduced peptide than on its oligomeric size. The results also suggest that the presence of the alpha-crystallin domain and hydrophobic clefts on the protein surface, which correlate poorly with oligomeric size, are important for chaperone function.
Saha
S
S
Protein Chemistry Laboratory, Department of Chemistry, Bose Institute, Kolkata, India.
Das
K P
KP
D000817:Anilino Naphthalenesulfonates / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007328:Insulin / Q000737:chemistry / Q000378:metabolism; D018832:Molecular Chaperones / Q000737:chemistry* / Q000378:metabolism*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D013329:Structure-Activity Relationship; D013696:Temperature; D014357:Trypsin / Q000378:metabolism*; D038202:alpha-Crystallin A Chain / Q000737:chemistry* / Q000378:metabolism*; D038203:alpha-Crystallin B Chain / Q000737:chemistry* / Q000378:metabolism*
D000817:Anilino Naphthalenesulfonates; D007328:Insulin; D018832:Molecular Chaperones; D010446:Peptide Fragments; D038202:alpha-Crystallin A Chain; D038203:alpha-Crystallin B Chain; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate); D014357:Trypsin