A calcium-insoluble 6.4 S protein derived from sea urchin cortical granule exudate.
A major protein component of the sea urchin, Strongylocentrotus purpuratus, cortical granule exudate has been purified and characterized. In the absence of divalent cations, the native, soluble protein has a sedimentation coefficient at infinite dilution of 6.4 S and a molecular weight from sedimentation equilibrium measurements of 2.8 +/- 0.3 X 10(5). These and other data indicate that the protein assumes an elongated, rod-like structure in solution. The protein is greater than 95% homogeneous as judged by agarose- and sodium dodecyl sulfate-gel electrophoresis. In the latter experiments, the protein shows a relative molecular weight of 1.8 X 10(5) and is clearly distinct from the 11.6 S protein described earlier which shows two bands corresponding to 3.2 X 10(5) and 2.1 X 10(5). The 6.4 S protein is the major protein of the calcium-insoluble fraction of cortical granule exudate and contributes to the formation of the extracellular investments of the sea urchin embryo. Using a light-scattering assay, we show that the purified protein retains the ability to aggregate in the presence of divalent cations mirroring its assembly in vivo. Calcium ion alone is able to initiate this reaction and the rate of precipitation increases with calcium concentration. Magnesium alone is ineffective in this regard but, in combination, the two ions act synergistically. Strontium and barium can substitute for calcium, but higher concentrations of the former cations are required to produce an equivalent effect.
Justice
R W
RW
Department of Biology, University of California, Riverside 92521.
Gottschling
C F
CF
Carroll
E J
EJ
Nagel
G M
GM
D000818:Animals; D002118:Calcium; D002458:Cell Fractionation; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D004527:Egg Proteins / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005122:Exudates and Transudates / Q000032:analysis*; D006818:Hyalin / Q000032:analysis*; D009391:Nephelometry and Turbidimetry; D012617:Sea Urchins; D012995:Solubility
D004527:Egg Proteins; D002118:Calcium