A protein of the basal lamina of the sea urchin embryo.
The purification, biochemical characterization and functional features of a novel extracellular matrix protein are described. This protein is a component of the basal lamina found in embryos from the sea urchin species Paracentrotus lividus and Hemicentrotus pulcherrimus. The protein has been named Pl-200K or Hp-200K, respectively, because of the species from which it was isolated and its apparent molecular weight in SDS-PAGE under reducing conditions. It has been purified from unfertilized eggs where it is found packed within cytoplasmic granules, and has different binding affinities to type I collagen and heparin, as assessed by affinity chromatography columns. By indirect immunofluorescence experiments it was shown that, upon fertilization, the protein becomes extracellular, polarized at the basal surface of ectoderm cells, and on the surface of primary mesenchyme cells at the blastula and gastrula stages. The protein serves as an adhesive substrate, as shown by an in vitro binding assay where cells dissociated from blastula embryos were settled on 200K protein-coated substrates. To examine the involvement of the protein in morphogenesis of sea urchin embryo, early blastula embryos were microinjected with anti-200K Fab fragments and further development was followed. When control embryos reached the pluteus stage, microinjected embryos showed severe abnormalities in arms and skeleton elongation and patterning. On the basis of current results, it was proposed that 200K protein is involved in the regulation of sea urchin embryo skeletogenesis.
Tesoro
V
V
Istituto di Biologia dello Sviluppo del Consiglio Nazionale delle Ricerche, Palermo, Italy.
Zito
F
F
Yokota
Y
Y
Nakano
E
E
Sciarrino
S
S
Matranga
V
V
D000818:Animals; D001485:Basement Membrane / Q000378:metabolism; D002448:Cell Adhesion; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D012617:Sea Urchins / Q000196:embryology*
D016326:Extracellular Matrix Proteins