Immunodetection of Xenopus bone morphogenetic protein-1 in adult and embryonic cells.
In order to analyze biochemical properties of Xenopus bone morphogenetic protein-1 (XBMP-1), rabbit antiserum (alpha-B1) was raised against a synthetic peptide (P1) corresponding to a hydrophilic N-terminal region. XBMP-1B (Xtld) synthesized in the reticulocyte lysate was successfully immunoprecipitated by this antiserum. This precipitation was completely blocked when P1 was added to the reaction, indicating that alpha-B1 recognized XBMP-1B specifically. In Western blot analysis, two distinct sizes of protein (107 and 34 kD) were detected in hind limbs in metamorphosing animals. Both proteins were detected in various adult tissues such as lung, liver, kidney, heart, muscle, intestine, brain, and testis. The mixing of the liver and muscle extracts, and the following detection of immunoreactive proteins suggested that the 34 kD band was a proteolytic product of the 107 kD protein. In the embryonic extracts from the unfertilized egg (stage 0) to swimming tadpoles (stage 40), a 63 kD protein was detected in addition to the 107 kD protein. We also showed that the 107 kD protein was much more expressed in the animal half of the unfertilized eggs than in the vegetal half, but that it was ubiquitously expressed in the gastrula embryos. We suggest that the 63 and 107 kD proteins correspond to full-length proteins encoded by XBMP-1A and XBMP-1B genes, and these proteins are expressed in embryo and in various adult tissues.
Ishimura
A
A
Department of Biology, Faculty of Science, Niigata University, Ikarashi, Japan.
Princler
G L
GL
Lin
J J
JJ
Kung
H F
HF
Maéno
M
M
D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D055395:Bone Morphogenetic Protein 1; D019485:Bone Morphogenetic Proteins; D007106:Immune Sera; D015151:Immunoblotting; D007814:Larva / Q000276:immunology; D008666:Metalloendopeptidases / Q000276:immunology* / Q000378:metabolism; D011233:Precipitin Tests; D013997:Time Factors; D014018:Tissue Distribution; D014981:Xenopus / Q000196:embryology* / Q000378:metabolism*; D029867:Xenopus Proteins
D000906:Antibodies; D019485:Bone Morphogenetic Proteins; D007106:Immune Sera; D029867:Xenopus Proteins; D008666:Metalloendopeptidases; C109187:TLL2 protein, Xenopus; D055395:Bone Morphogenetic Protein 1