A peptide sequence-YSGVCHTDLHAWHGDWPLPVK [40-60]-in yeast alcohol dehydrogenase prevents the aggregation of denatured substrate proteins.
The structural and functional characteristics of a yeast alcohol dehydrogenase (ADH) peptide (YSGVCHTDLHAWHGDWPLPVK, residues 40-60) have been studied in detail. The peptide is hydrophobic in nature, binds the hydrophobic probe bis-ANS, and is mostly present in a random coil conformation. It shows chaperone-like activity by preventing dithiothreitol (DTT)-induced aggregation of insulin at 27 degrees C, oxidation-induced aggregation of gamma-crystallin at 37 degrees C, and aggregation of thermally denatured ADH and beta(L)-crystallins at 52 degrees C. However, the ADH peptide does not solubilize protein aggregates as do surfactants. Substitution of Pro for His in the ADH peptide leads to diminished anti-aggregation activity. Further, analysis of ADH incubated at 47 degrees C suggests that a significant portion of the enzyme remains as soluble inactive protein with negligible conformational change. Therefore, we propose that the residues 40-60 in native protein may be an intramolecular chaperone site of yeast ADH.
Bhattacharyya
Jaya
J
Mason Eye Institute, Departments of Ophthalmology and Biochemistry, University of Missouri, Columbia, MO 65212, USA.
Santhoshkumar
P
P
Sharma
K Krishna
KK
D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000817:Anilino Naphthalenesulfonates / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D003432:Cross-Linking Reagents / Q000378:metabolism; D004229:Dithiothreitol / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D007328:Insulin / Q000378:metabolism; D018832:Molecular Chaperones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D013696:Temperature; D015003:Yeasts / Q000201:enzymology* / Q000235:genetics; D038204:beta-Crystallins / Q000378:metabolism; D038222:gamma-Crystallins / Q000378:metabolism
D000817:Anilino Naphthalenesulfonates; D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D007328:Insulin; D018832:Molecular Chaperones; D010446:Peptide Fragments; D038204:beta-Crystallins; D038222:gamma-Crystallins; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate); D000426:Alcohol Dehydrogenase; D004229:Dithiothreitol