Does muscle-type myosin have ADPase activity?

Adenosine diphosphate (ADP) is a nucleotide that is structurally very similar to ATP but lacks one of the two high-energy bonds due to hydrolysis. In muscle studies, ADP is usually considered exclusively as a product formed during myosin cross-bridge cycling and is not otherwise involved in this process. In our study, we question the widely held view of ADP as a final product formed during muscle contraction. Using biophysical and biochemical methods, we managed to show that ADP can act as a substrate for myosins in at least three types of muscles: smooth and striated adductor muscles of bivalves (Mytilidae and Pectinidae), and also vertebrate skeletal muscles. According to our data, the differences in the effect of ATP and ADP on the optical, biochemical, and structural properties of actomyosins are exclusively quantitative. We explain the previous ideas about ADP as a compound capable of inhibiting the ATPase activity of actomyosin by the ability of ATP and ADP to depolymerize the polymeric myosin when the concentration in the medium reaches more than 0.3 mM.

A Preparative Method for the Isolation of Calponin from Molluscan Catch Muscle.

We describe the development of a preparative method to isolate molluscan catch muscle, calponin. This method is based on the ability of calponin to interact with actin in a temperature-dependent manner. After extracting thin filaments, as previously described, the extract was ultracentrifuged at 2 °C. While other surface proteins of thin filaments co-precipitated with actin, calponin, along with some minor contaminants, remained in the supernatant. Calponin was purified through cation-exchange chromatography. The yield of pure protein was four-fold higher than that achieved through high-temperature extraction. To evaluate functionally isolated proteins, we determined the effect of calponin on Mg2+-ATPase activity of hybrid and non-hybrid actomyosin. The degree of ATPase inhibition was consistent with previously published data but strongly dependent on the environmental conditions and source of actin and myosin used. Furthermore, at low concentrations, calponin could induce the ATPase activity of hybrid actomyosin. This result was consistent with data indicating that calponin can modulate actin conformation to increase the relative content of "switched on" actin monomers in thin filaments. We assume that calponin obtained by the isolation method proposed herein is a fully functional protein that can both inhibit and induce the ATPase activity.

Are myoepithelial cells confined to genital coelomic sinus in the gonads of sea stars?

An ultrastructural study of the gonadal wall in 10 sea star species from the orders Forcipulatida, Paxillosida, Spinulosida, Valvatida and Velatida has shown variations in the presence of myoepithelial cells in the visceral peritoneal epithelium. These cells have only been found in the peritoneal epithelium of the gonads in Aphelasterias japonica (Forcipulatida: Asteriidae), Asterias amurensis (Forcipulatida: Asteriidae), Distolasterias nipon (Forcipulatida: Asteriidae), Diplopteraster multipes (Velatida: Pterasteridae), Luidia quinaria (Paxillosida: Ctenodiscidae), and Pteraster sp. (Velatida: Pterasteridae). Our results may shed light on the evolution of peritoneal epithelium of sea star gonads. It is probable that, initially sea stars had myoepithelial cells in visceral peritoneal epithelium of the gonads. The species from the orders Forcipulatida and Velatida have retained this plesiomorphic state, while many species from the orders Paxillosida, Spinulosida and Valvatida have lost myoepithelial cells from visceral peritoneal epithelium of their gonads.

Hematopoiesis in Bivalvia larvae: Cellular origin, differentiation of hemocytes, and neoplasia.

Hemocytes play vital roles in the immune response. Despite progress in the characterization of molluscan hemocytes and immune cells, including their cellular receptors and signal transduction pathways, the processes that lead to their differentiation in bivalve larvae remain unknown. Furthermore, the molecular mechanisms of that decide hemocyte stem cell fate and self-renewal during development remain poorly characterized. Similar to adult mollusks, the larvae are filter feeders and are highly susceptible to pathogens and biotoxins; therefore, it is important to understand the development and function of their immune system. This review summarizes the current data on the appearance of elements of the immune system in bivalve larvae. I have discussed why the immune cells emerge before the circular blood vessel system, which differentiates at the late stages of development. I also discuss how molluscan hemocytes are involved in the development of disseminated neoplasia.

Molluscan catch muscle myorod and its N-terminal peptide bind to F-actin and myosin in a phosphorylation-dependent manner.

Myorod is expressed exclusively in molluscan catch muscle and localizes on the surface of thick filaments together with twitchin and myosin. This protein is an alternatively spliced product of the myosin heavy-chain gene containing the C-terminal rod part of myosin and a unique N-terminal domain. We have recently reported that this unique domain is a target for phosphorylation by gizzard smooth muscle myosin light chain kinase (MLCK) and molluscan twitchin, which contains a MLCK-like domain. To elucidate the role of myorod phosphorylation in catch muscle, a peptide corresponding to the specific N-terminal region of the protein was synthesized in phosphorylated and unphosphorylated form. We report, for the first time, that unphosphorylated full-length myorod and its unphosphorylated N-terminal synthetic peptide are able to interact with rabbit F-actin and thin filaments from molluscan catch muscle. The binding between thin filaments and the peptide was Ca²+-dependent. In addition, we found that phosphorylated N-terminal peptide of myorod has higher affinity for myosin compared to the unphosphorylated peptide. Together, these observations suggest the direct involvement of the N-terminal domain of myorod in the regulation of molluscan catch muscle.