Biomedical subjects

Masaru Kawamura

Publications and source records attributed to Masaru Kawamura.

5 recordsLinked to original sources

PubMed2005

Involvement in K+ access of Leu318 at the extracellular domain flanking M3 and M4 of the Na+,K+-ATPase alpha-subunit.

The effect of point mutation in the sequence 316TWLE319, which occurs in the extracellular loop flanking the third (M3) and the fourth (M4) transmembrane segment (L3/4) of the Na+,K+-ATPase alpha-subunit, was examined. Mutation of Glu319 to Asp yielded an enzyme with full activity, whereas substituting Glu319 to Ala resulted in a severe loss of activity. A negative charge was introduced along the sequence, one residue at a time, from Thr316 to Leu318 (by E-scanning) in the mutant construct with Glu319 already mutated to Gln. The activity that had been reduced to 60% by the mutation of Glu319 to Gln was restored upon the introduction of a negative charge by E-scanning. When Leu318 was replaced by Glu in a series of scanning experiments, the K+ sensitivity of the ATPase activity was lowered. The lowering of K+ sensitivity was further demonstrated when a mutation of Leu318 to Glu was introduced into the wild-type enzyme. Furthermore, mutants with Leu318 to Gln, Arg, and Phe displayed lower K+ sensitivity similar to that of Leu318 to Glu mutant. Leu318 may be in access path for K+, and any substitution at this position may interfere with access of K+ from outside the cell.

Animals↗

PubMed2005

Fe(II)/Cu(I)-dependent P-type ATPase activity in the liver of Long-Evans cinnamon rats.

This study examined Fe(II)-dependent ATPase activity in OTG (octylthioglucoside) -treated microsomes isolated from Wistar and LEC rats. The ATPase activity of the liver OTG-microsomes from Wistar rats increased sharply in the 5-150 microM range of Fe(II) with a K0.5 value of 23.9+/-3.6 microM, while the activity of LEC rat liver microsomes increased with increasing Fe(II) up to 500 microM with a K0.5 value of 64.4+/-8.1 microM. The K0.5 values for Fe(II)-dependent ATPase activity of spleen OTG-microsomes were nearly identical at 59.3 microM in the Wistar rat and 63.7 microM in the LEC rats with a similar level of activity at each Fe(II) concentration in both strains of animals. These results indicated that there are two types of Fe(II)-dependent ATPase with different Fe(II) sensitivity, a high sensitive (H) and a low sensitive (L) type, and that the H-type activity was specific to the liver. The H-type activity was, however, deficient in the liver of LEC rats that accumulate copper and iron in hepatocytes as a result of mutations in the Wilson's disease protein (WNDP). On the basis of these results, together with the similarity in optimal conditions required for full activity of the enzyme, we conclude that the Fe(II)-dependent ATPase (H-type) and WNDP may be identical.

Adenosine Triphosphatases↗

PubMed2003

Transient association of the sarcoplasmic reticulum Ca2+ ATPase with the Na+/K+-ATPase and H+/K+-ATPase beta-subunits during its biogenesis in Xenopus oocytes.

We examined the effect of the beta-subunits of the Na+/K+ and H+/K+ ATPases on the biogenesis of the sarcoplasmic reticulum (SR) Ca2+ ATPase in Xenopus oocytes. Oocytes were simultaneously injected with cRNAs for both the SR Ca2+ ATPase and the beta-subunit of the Na+/K+ or the H+/K+ ATPase. Immunoprecipitation with antiserum specific for the beta-subunit of the Na+/K+ or the H+/K+ ATPase yielded not only the respective beta-subunit but also the SR Ca2+ ATPase, indicating that the SR Ca2+ ATPase was associated with the beta-subunits of the Na+/K+ and the H+/K+ ATPases. Pulse-chase experiments revealed that the complex between the SR Ca2+ ATPase and the beta-subunit of the Na+/K+ ATPase was formed transiently and dissociated during the course of maturation. This is the first report that demonstrates the association of the SR Ca2+ ATPase with the beta-subunit of the Na+/K+ and H+/K+ ATPases.

Adenosine Triphosphatases↗

PubMed2003

Functional role of the N-terminus of Na(+),K(+)-ATPase alpha-subunit as an inactivation gate of palytoxin-induced pump channel.

The N-terminus of the Na(+),K(+)-ATPase alpha-subunit shows some homology to that of Shaker-B K(+) channels; the latter has been shown to mediate the N-type channel inactivation in a ball-and-chain mechanism. When the Torpedo Na(+),K(+)-ATPase is expressed in Xenopus oocytes and the pump is transformed into an ion channel with palytoxin (PTX), the channel exhibits a time-dependent inactivation gating at positive potentials. The inactivation gating is eliminated when the N-terminus is truncated by deleting the first 35 amino acids after the initial methionine. The inactivation gating is restored when a synthetic N-terminal peptide is applied to the truncated pumps at the intracellular surface. Truncated pumps generate no electrogenic current and exhibit an altered stoichiometry for active transport. Thus, the N-terminus of the alpha-subunit appears to act like an inactivation gate and performs a critical step in the Na(+),K(+)-ATPase pumping function.

Acrylamides↗

PubMed2002

Monomeric sarcoplasmic reticulum Ca(2+)-ATPase is functionally active.

cRNAs for wild type and a functionless mutant of sarcoplasmic reticulum (SR) Ca(2+)-ATPase were co-injected into Xenopus oocytes in various molar ratios. SR Ca(2+)-ATPase activity of the microsomal fraction prepared from the injected oocytes increased linearly with increasing amounts of wild type cRNA. This result suggests that monomers are functional units of membrane-bound SR Ca(2+)-ATPase.

Animals↗