Oleosins are unique and major proteins localized on the surface of oil bodies in diverse seed species. We purified five different KD 20), and raised chicken antibodies against them. These antibodies were used to test for immunological cross-reactivity among oleosins from diverse seed species. Within the same seed species, antibodies raised against one oleosin isoform did not cross-react with the other oleosin isoform (i.e. between maize oleosins KD 16 and KD 18, and between soybean oleosins KD 18 and KD 24). However, the respective antibodies were able to recognize oleosins from other seed species. Where interspecies cross-reactivity occurred, the results suggest that there are at least two immunologically distinct isoforms of oleosins present in diverse seed species, one of lower Mr, and another one of higher Mr. This suggestion is also supported by the relative similarities between the amino acid sequence of a small portion of rapeseed oleosin KD 20 and those of maize oleosins KD 16 and KD 18. In maize kemel, there was a tissue-specific differential presentation of the three oleosins, KD 16, KD 18, and KD 19, in the oil-storing scutellum, embryonic axis, and aleurone layer. The phylogenetic relationship between the high and low Mr isoforms within the same, and among diverse, seed species is discussed.

The structural characteristics of myelin basic protein (MBP) involved in protein-protein and protein-lipid interactions were investigated. Rabbit MBP could bind calmodulin (CaM) in the presence of Ca2+ to form a complex that remained undissociated in 8 M urea. However, no tight complex formation was observed when the divalent cation was absent. These results suggest that MBP may contain a hydrophobic domain similar to those in the other well-characterized CaM-binding proteins. The stoichiometry of calmodulin binding to MBP was approximately 1:1. Prior limited proteolysis of MBP with trypsin abolished the formation of the MBP-CaM complex, indicating that the entire MBP polypeptide may be involved in the recognition of the hydrophobic clefts in CaM. MBP also formed tight complexes with gangliosides, but the presence of Ca2+ was not required. Binding of gangliosides to MBP-CaM complex released CaM from the complex. The ganglioside-binding sites in MBP were determined after trisecting the protein at two glutamic acid residues with Staphylococcus aureus V8 protease. Subsequent binding studies revealed that a 9.5-kDa polypeptide, which may correspond to the NH2-terminal domain (residues 1-83) of MBP, had higher affinity for the binding of lucifer yellow CH-labeled GM1 than did the other two polypeptides, of apparent molecular mass (Mr) 5,500 and 4,500, respectively. Among the various proteins in purified guinea pig brain myelin, synaptosomes, and synaptosomal membranes, MBP was found to have the highest affinity in binding lucifer yellow CH-GM1.

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