Induction of either contractile or structural actin-based gels in sea urchin egg cytoplasmic extract.
The gel formed by warming the 100,000 g supernate of isotonic extracts of sea urchin eggs to 40 degrees C is made up of actin and two additional proteins of mol wt of 58,000 and 220,000. Actin and 58,000 form a characteristic structural unit which has now been identified in the microvilli of the urchin egg and in the filopods of urchin coelomocytes. However, egg extract gels did not contract as those from other cell types do, and the aim of these experiments was to determine the reason for this lack of contraction. Although the extracts are dialyzed to a low ionic strength, myosin is present in soluble form and makes up approximately 1% of the protein of the extract. It becomes insoluble in the presence of high ATP concentrations at 0 degrees C, and the precipitate formed under these conditions consists almost entirely of myosin. This procedure provides a simple method of isolating relatively pure myosin without affecting other extract components and functions. Contraction will follow gelation in these extracts if the temperature and time of incubation used to induce actin polymerization are reduced to minimize myosin inactivation. At the optimal ATP and KCl concentration for contraction, the contracted material has an additional 250,000 component and contains very little 58,000. The conditions found to provide maximum gel yields favor the formation of the actin-58,000-220,000 structural gel, while reduced temperature and increase in KCl concentration results in a contractile gel whose composition is similar to those reported from amoeboid cell types. Both the structural protein cores found in the egg microvilli and a gel contraction related to the amoeboid motion which is seen in later urchin embryonic development can thus be induced in vitro in the same extract.
D000199:Actins / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002465:Cell Movement; D003599:Cytoskeleton / Q000648:ultrastructure*; D005260:Female; D005782:Gels; D008274:Magnesium / Q000378:metabolism; D008970:Molecular Weight; D009218:Myosins / Q000378:metabolism; D010063:Ovum / Q000648:ultrastructure*; D011189:Potassium Chloride / Q000378:metabolism; D012617:Sea Urchins / Q000648:ultrastructure*; D013696:Temperature
D000199:Actins; D002352:Carrier Proteins; D005782:Gels; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D009218:Myosins; D008274:Magnesium