. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. CHARACTERIZATION OF CORAL MORPHOGEN 113 loop. 4 6 Hydrolysis (hr) Figure 9. Time-course of solubilization of morphogen by mild al- kaline hydrolysis. The decalcified, cell wall-associated inducer from Hy- drolithtm huert>csenii was incubated with vigorous stirring at pH At the times indicated, aliquots were withdrawn, neutralized to pH filtered through nitrocellulose filters ( ^m. 47 mm diam.), and the soluble fractions assayed (in duplicate 10 ml samples) under the standard conditions. Details are in Materi
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. CHARACTERIZATION OF CORAL MORPHOGEN 113 loop. 4 6 Hydrolysis (hr) Figure 9. Time-course of solubilization of morphogen by mild al- kaline hydrolysis. The decalcified, cell wall-associated inducer from Hy- drolithtm huert>csenii was incubated with vigorous stirring at pH At the times indicated, aliquots were withdrawn, neutralized to pH filtered through nitrocellulose filters ( ^m. 47 mm diam.), and the soluble fractions assayed (in duplicate 10 ml samples) under the standard conditions. Details are in Materials and Methods. Results shown are the average percentages of larvae induced to attach and metamorphose by the soluble fractions, ± Results of a parallel and otherwise identical incubation held continuously at pH are included for comparison. polysaccharide (cleaved by hyaluronidase, agarase, and sulfatase), with multiple, substituted residues of N-ace- tylglucosamine (linkages cleaved by lysozyme and endo- itf-galactosidase) and multiple residues of galactose (link- ages cleaved by agarase and endo-/3-galactosidase). Protein (or peptide) does not appear to be an essential component, nor do several specific free or terminal sugars or saccharide units. Hydrolysis with dilute alkali releases small anionic morphogen containing sulfate The finding that specific endopolysaccharidases can re- lease a soluble morphogen from the insoluble cell wall- associated fraction suggested that non-enzymatic, partial hydrolysis under mild conditions might yield similar re- sults. While some desulfation also would occur, hydrolysis with sufficiently dilute alkali would be expected to yield oligosaccharide fragments with retention of at least some of the original sulfate groups (Percival and McDowell, 1967). As predicted, exposure of the decalcified, partic- ulate cell wall fraction to dilute alkali (pH 11-12) released a soluble, sulfate-containing morphogen. The increase in yield of solubl
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