Hyalin, a sea urchin extraembryonic matrix protein: relationship between calcium binding and hyalin gelation.
The protein hyalin, a major component of the sea urchin extraembryonic hyaline layer, was previously shown to undergo a Ca(2+)-induced self-association into large aggregates (gelation). This reaction represented a major step in assembly of the layer. In the experiments reported here, digestion with trypsin resulted in a rapid dissociation of hyalin into a mixture of peptides which retained the capacity to bind Ca2+. However, unlike intact hyalin, none of these peptides associated into large aggregates (gelation) in the presence of Ca2+, Mg2+, and NaCl. Loss of the ability to undergo gelation was not accompanied by any significant change in the content of acidic plus amide amino acid residues. Decreasing the pH to 5.6 resulted in a loss of 25% of hyalin's Ca(2+)-binding capacity but had no effect on the ability of the protein to undergo gelation. Peptide fragments were only partially effective at inhibiting hyalin gelation. Clearly, not all the Ca(2+)-binding sites were required for hyalin gelation and Ca2+ binding alone was insufficient to drive this reaction. In addition, hyalin appeared to possess two classes of protein-protein interaction domains, one of which was essential for gelation.
Robinson
J J
JJ
Department of Biochemistry, Memorial University of Newfoundland, St. John's, Canada.
Hall
D
D
Brennan
C
C
Kean
P
P
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000737:chemistry* / Q000378:metabolism*; D005782:Gels; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry; D012617:Sea Urchins / Q000196:embryology*; D013329:Structure-Activity Relationship
D000596:Amino Acids; D002135:Calcium-Binding Proteins; D016326:Extracellular Matrix Proteins; D005782:Gels; D010446:Peptide Fragments; C076585:hyalin protein, Echinoidea; D002118:Calcium