0000000000082422
AUTHOR
Donatella Ciulla
Studies on (echinodermata) coelomocyte lysate I. Hemolytic activity of coelomocyte hemolysins
Abstract The Holothuria polii coelomocyte lysate contains two trypsin-resistant lytic proteins having different chemico-physical properties : a calcium dependent and heat-labile hemolysin that is probably a constitutive component of the coelomic fluid, and another calcium independent and heat-stable one that is released after immunological stimulation; it is therefore not detectable in natural conditions. The sphingomyelin seems to be the membrane receptor with which both hemolysins interact producing lysis.
The hemolysin-producer coelomocytes in Holothuria polii
Using sodium metrizoate discontinuous gradients, two hemolysin-producer amebocyte populations have been separated from total circulating Holothuria polii coelomocytes. The amebocytes of population 1 are responsible for the production of the calcium-dependent and temperature-labile hemolysin, whereas those of population 2 produce the calcium-independent and temperature-stable one. The intracytoplasmic hemolysins were evidenced also by immunofluorescence. Petaloid and filipodial amebocytes were the only positive cell types.
Studies on Holothuriapolii (echinodermata) coelomocyte lysate II. Isolation of coelomocyte hemolysins
The lytic activity of the Holothuria polii coelomocyte lysate resides in two electrophoretically distinct hemolysins identified as He1 and He2. He1 represents the calcium dependent, heat-labile component whereas He2 is calcium independent and heat-stable. The two hemolysins share serological identity. Both hemolysins appear as single protein molecules of 80KDa molecular weight by SDS-PAGE and transblotting analysis under non-reducing conditions. However under reducing conditions, they are doublets of 76 and 80KDa molecular weight. The hypothesis that the two hemolysins could be isoforms is discussed.