Molluscan hemocyanin, a copper-containing oxygen transporter, is one of the largest known proteins. Although molluscan hemocyanins are currently applied as immunotherapeutic agents, their precise structure has not been determined because of their enormous size. Here, we have determined the first X-ray crystal structure of intact molluscan hemocyanin. The structure unveiled the architecture of the 3.8-MDa supermolecule composed of homologous functional units (FUs), wherein the dimers of FUs hierarchically associated to form the entire cylindrical decamer. Most of the specific inter-FU interactions were localized at narrow regions in the FU dimers, suggesting that rigid FU dimers formed by specific interactions assemble with flexibility. Furthermore, the roles of carbohydrates in assembly and allosteric effect, and conserved sulfur-containing residues in copper incorporation, were revealed. The precise structural information obtained in this study will accelerate our understanding of the molecular basis of hemocyanin and its future applications.

Jumbo squid was very similar to Japanese common squid in terms of myofibrillar Ca2+‐, Mg2+‐ and K+(EDTA)‐ATPase activities. Myofibrils of jumbo squid were significantly stabilized upon addition of Ca2+ and destabilized by increasing KCl concentration for heating. Incubation of muscle homogenate of jumbo squid induced a selective cleavage of myosin into two major fragments and the cleavage was inhibited by EDTA. Autolysis was prominent at and above 0.3 M NaCl where myosin filaments dissolve. The enzyme involved in the autolysis was proved to be unstable showing maximal autolysis rate at 25°C. Washing the homogenate partially reduced the autolysis activity.

Thermal gelation of salted squid mantle muscle paste was studied in relation to endogenous proteases and transglutaminase. Myosin in the paste was preferentially degraded into 130-kDa and 90-kDa fragments at an optimum temperature of 30°C. Degradation was inhibited with EDTA or 1,10-phenanthroline, suggesting the presence of metalloproteases. Myosin degradation was markedly reduced above 40°C. Although 10 mM Ca 2+ increased cross-linking of myosin heavy chains by activating the endogenous transglutaminase, setting effect on thermal gelation of the paste was offset by degradation induced by simultaneously activated calpains. Ca 2+ and the calpain inhibitor, E64, significantly improved the breaking strength and strain of thermal gels preincubated at 40°C.

The 2004 Mid Niigata Prefecture earthquake sequence produced a nearly 1-km-long, N-S to NNW-SSEstriking, west-side-up surface rupture along a previously unmapped fault at Obiro, Uonuma City, eastern margin of the epicentral region. The maximum vertical displacement is 20 cm. The topographic and geometric features of the surface rupture are indicative of reverse faulting with dip to the west, which is consistent with focal mechanism solutions and aftershock distribution. Along the major active faults and folds that form the framework of landforms in the epicentral region, systematic surface ruptures were not found. Exceptionally small rupture dimension and offset amount of the surface rupture compared with those of the modeled fault suggest that the fault slip appears to have been accommodated by both internal deformation in thick Neogene-Quaternary deposits and growth of the folds and blind thrusts around the epicentral region.