Partial amino acid sequence analysis of rabbit fast-twitch skeletal muscle calsequestrin permitted the construction synthetic oligonucleotides that were used as both primers and probes for synthesis isolation cDNAs encoding from neonatal libraries. The cDNA encodes a processed protein 367 residues with Mr 42,435 28-residue amino-terminal signal sequence. deduced agreed closely portions mature sequenced using standard sequencing. protein, however, contains an acidic carboxyl-terminal...

Protein phosphatase 2C (PP2C) is implicated in the negative regulation of stress-activated protein kinase cascades yeast and mammalian cells. In this study, we determined role PP2Cβ-1, a major isoform PP2C, TAK1 signaling pathway, cascade that activated by interleukin-1, transforming growth factor-β, or stress. Ectopic expression PP2Cβ-1 inhibited TAK1-mediated mitogen-activated 4-c-Jun amino-terminal 6-p38 pathways. vitro, dephosphorylated inactivated TAK1. Coimmunoprecipitation experiments...

Although quercetin has suppressed bone resorption in several animal studies, its target cells and the mechanism of action related to not been fully elucidated. We investigated effect on differentiation activation osteoclasts. used cocultures mouse spleen ST2 cells, cultures osteoclast progenitor {M-CSF-dependent (MD) from marrow murine monocytic RAW 264 (RAW) cells}. Quercetin dose-dependently inhibited osteoclast-like (OCL) cell formation at 2—5 μM concentration both coculture MD culture....

Protein phosphatase 2Cα (PP2Cα) or PP2Cβ‐1 expressed in COS7 cells suppressed anisomycin‐ and NaCl‐enhanced phosphorylations of p38 co‐expressed the cells. PP2Cα expression also both basal stress‐enhanced MKK3b MKK6b, which are upstream protein kinases p38, MKK4, is one major JNK. Basal activity MKK7, another kinase JNK, was by expression. However, as well serum‐activated phosphorylation MKK1a, an ERKs, not affected PP2Cβ PP2Cβ‐1. A catalytically inactive mutant further enhanced...

Sanguinarine, a plant alkaloid, was identified as potent and specific protein phosphatase (PP) 2C inhibitor. It inhibited PP2C competitively with respect to α-casein (Ki=0.68 μm) showed selectivity for compared PP1, PP2A, PP2B in vitro. In vivo, sanguinarine cytotoxicity toward human promyelocytic leukemia cell line HL60, an IC50 value of 0.37 μm, induced apoptosis through caspase-3/7-dependent mechanism involving the phosphorylation p38, PP2Cα substrate. The activity by partially p38...

Background/Objectives: Facial bone density, including the jawbone, declines earlier than that of lumbar spine and calcaneus. Calcium maltobionate is reported to mitigate resorption maintain density in post-menopausal women, but its effects on facial remain understudied. Therefore, this study compared variations mineral with calcaneal markers among healthy examining differences between pre- post-menopause continuous calcium intake. Methods: This randomized, double-blind, placebo-controlled,...

Type I allergy is a hypersensitivity reaction that occurs as result of antigen stimulation.Allergic disorders such asthma, rhinitis, atopic dermatitis, and anaphylactic shock are classifi ed type allergies.The number patients with these allergies has been increasing in most parts the world (1, 2).Present allergic medicines suppress symptoms but do not cure illness completely (3,4).Consequently, have to continue medication long exist.Furthermore, medications side effects drowsiness (5).Thus,...

During activation of platelets by thrombin phosphorylation Thr(558) in the C-terminal domain membrane-F-actin linking protein moesin increases transiently, and this correlates with protrusion filopodial structures. Calyculin A enhances inhibition phosphatases. To measure moesin-specific activity, a nonradioactive enzyme-linked immunosorbent assay method was developed synthetic peptide Cys-Lys(555)-Tyr-Lys-Thr(P)-Leu-Arg(560) coupled to bovine serum albumin as substrate state-specific...

The structures of five distinct isoforms mammalian protein phosphatase 2Cβ (PP2Cβ-1, -2, -3, -4 and -5) have previously been found to differ only at their C-terminal regions. In the present study, we performed mutational analysis recombinant mouse PP2Cβ-1 determine functional domains molecule elucidate biochemical significance structural differences in isoforms. Differences affinity for [32P]phosphohistone but not [32P]phosphocasein were observed among PP2Cβ Deletion 12 amino acids from end,...

P19 embryonal carcinoma cells are known to differentiate into neurons and glia when treated with relatively high concentrations (>100 nM) of retinoic acid (RA). Concomitant this RA‐induced neural differentiation, we observed an activation the c‐Jun amino‐terminal kinase (JNK). JNK was required for because dominant‐negative blocked differentiation. Studies using protein phosphatase inhibitors suggested that both okadaic acid‐sensitive phosphatase(s) C participate in JNK.

The ubiquitin/proteasome system plays significant and important roles in the regulation of metabolism various proteins. dysfunction this is involved several diseases, for example, cancer, neurogenic diseases chronic inflammation. Therefore, compounds, which regulate system, might be candidates development use as clinical drugs. Saccharomyces cerevisiae mutant (rsp5(A401E)) has a single amino acid change, Ala401Glu, RSP5 gene, encodes an essential E3 ubiquitin ligase, hypersensitive to...

Abstract Background Osteoporosis is a debilitating disease characterized by decreased bone density. In this study, we evaluated the anti‐osteoporotic effect of maltobionic acid (MB), one components honey, and calcium maltobionate (MBCa) on density metabolism using mouse model osteoporosis. The underlying mechanisms MB MBCa action were also investigated. Results Ovariectomized (OVX) mice fed diet containing or for 80 days, femoral mineral content (BMD) evaluated. As expected, OVX reduced BMD;...

Mouse P19 embryonal carcinoma cells in aggregation culture the presence of 10−6M retinoic acid followed by monolayer differentiate into nerve and glial cells. In this study, we demonstrated that neurofilament-L (NF-L) mRNA protein levels these were enhanced accordance with their acid-induced neural differentiation. Okadaic (OA) treatment markedly suppressed differentiation-dependent NF-L gene expression increase neurite outgrowth Similar results obtained when tautomycin was used instead OA,...

We examined the in vitro digestibility of maltobionic acid, obtained from enzymatic oxidation maltose, its utilization by intestinal bacteria, and biological effects on bowel movements healthy subjects. found that acid is not digested saliva, gastric juice, or pancreatic juice. Moreover, it only to a small extent enzymes. Among 24 strains was selectively utilized Bifidobacterium dentium Bi. adolescentis. also evaluated influence long-term ingestion calcium salt Japanese women randomized,...

In this study we have cloned a novel member of mouse protein phosphatase 2C family, PP2Cζ, which is composed 507 amino acids and has unique N‐terminal region. The overall similarity the acid sequence between PP2Cζ PP2Cα was 22%. On Northern blot analysis found to be expressed specifically in testicular germ cells. COS7 cells able associate with ubiquitin conjugating enzyme 9 (UBC9) association enhanced by co‐expression small ubiquitin‐related modifier‐1 (SUMO‐1), suggesting that exhibits its...

Abstract Maltobionic acid is known to have an inhibitory effect on the differentiation of osteoclasts, and it has also been reported in intervention trial that ingestion corn syrup solids containing maltobionic maintained increased bone density postmenopausal women. However, there no information whether improves metabolism humans. Therefore, we evaluated influence (maltobionic calcium salt) resorption markers healthy Japanese Forty‐one individuals were selected from 68 participants assigned...

In the course of our screening program for a new inhibitor Ca(2+) signalling pathway using mutant yeast [Saccharomyces cerevisiae (zds1Δ erg3Δ pdr1Δ pdr3Δ)], mouse PP2Cα activator, pisiferdiol, isolated from Chamaecyparis pisifera, was found to alleviate signal-mediated growth inhibition. Pisiferdiol showed inhibition activity against mpk1Δ strain compared with cnb1Δ and induced Li(+) sensitivity wild-type strain, indicating that it suppresses calcineurin in yeast. However, ptc1Δ by...