The effect of pH on the structure of recombinant chicken Hsp24, human Hsp27 and their 3D mutants mimicking phosphorylation ;at Ser15, Ser77/78, and Ser81/82 was analyzed. Circular dichroism and fluorescent spectroscopy indicate that changes of pH in the range 6.0-7.5 weakly affected the secondary and tertiary structure of the wild type proteins, but induced noticeable changes in the structure of their 3D mutants. According to size-exclusion chromatography and analytical ultracentrifugation variation of pH-induced pronounced changes in the quaternary structure of small heat shock proteins and acidification resulted in accumulation of large oligomers of Hsp24/27. It is concluded that small changes of pH strongly affect the quaternary structure of small heat shock proteins and by this means can influence their functioning in the cell. (C) 2004 Elsevier Inc. All rights reserved.