Cofilin is a widely distributed actin-modulating protein that has the ability to bind along side of F-actin and depolymerize in pH-dependent manner. We found phosphatidylinositol (PI), 4-monophosphate (PIP), 4,5-bisphosphate (PIP2) inhibited both actions cofilin dose-dependent manner, while inositol 1,4,5-triphosphate (IP3), 1-oleoyl-2-acetylglycerol (OAG), phosphatidylserine (PS), or phosphatidylcholine (PC) had little no effect on them. Gel filtration analyses showed PIP2 bound thereby...

Incubation of cultured cells under specific conditions induces a dramatic change in the actin organization: induction intranuclear and/or cytoplasmic rods (actin paracrystal-like intracellular structures). We have found that cofilin, 21-kDa actin-binding protein, is component these rods. Antibodies directed against cofilin labeled induced treated with dimethyl sulfoxide or exposed to heat shock and also incubated salt buffers. Moreover, we are not stained fluorescent phalloidin derivatives...

1984) Biochemistry 23,5307-5313), was here shown to be capable of reversibly controlling actin polymerization and depolymerization in a pH-sensitive manner.When cofilin reacted with F-actin at different pH, the depolymerized concentration (= monomeric concentration) higher elevated pH.At pH c 7.3, concentrations did not exceed -1 FM even presence excess amounts cofilin, whereas > 7.3 it increased proportion

Cofilin is a widely distributed actin-modulating protein that has abilities to bind along the side of F-actin and depolymerize F-actin. Both cofilin can be inhibited by phosphoinositides such as phosphatidylinositol, phosphatidylinositol 4-monophosphate, 4,5-bisphosphate (PIP2). We have previously shown synthetic dodecapeptide corresponding Trp104-Met115 potent inhibitor actin polymerization (Yonezawa, N., Nishida, E., Iida, K., Kumagai, H., Yahara, I., Sakai, H. (1991) J. Biol. Chem. 266,...

Cofilin is a widely distributed, pH-sensitive, actin-modulating protein with an apparent molecular mass of 21 kDa, which forms intranuclear and/or cytoplasmic actin/cofilin rods in cultured fibroblastic cells under specific conditions. In this study, cDNA library from porcine brain mRNA was constructed, and full-length cofilin clones were isolated by screening oligonucleotide probes. The deduced amino acid sequence 166 residues long contains Lys-Lys-Arg-Lys-Lys very similar to the nuclear...

Destrin is a mammalian 19-kDa protein that rapidly depolymerizes F-actin in stoichiometric manner. In this study, we isolated cDNA clones coding for destrin from porcine brain library. The deduced amino acid sequence of 165 residues long and very similar (71% identical) to cofilin, widely distributed, pH-sensitive actin-modulating protein. contains nearly identical with the putative nuclear transport signal cofilin hexapeptide amino-terminal (residues 2-7) tropomyosin, which shown be...

The sperm receptor activity of pig zona pellucida has been previously shown to exist in one the components, protein 3α (PZP3α), that can be purified after removal sialylated and/or sulfated N ‐acetylpoly(lactosamine) by digestion with endo‐β‐galactosidase. In this study, we examined whether N‐linked or O‐linked carbohydrate chains are involved pellucida. elimination from endo‐β‐galactosidase‐digested PZP3α ‐glycanase markedly reduced its inhibitory effect on sperm‐egg binding an vitro...

Cofilin and destrin are two related low molecular weight mammalian actin-binding proteins. is an F-actin side-binding pH-dependent actin-depolymerizing protein, a pH-independent protein. We have introduced few point mutations within sequence of cofilin. Biochemical analyses these mutant proteins clearly shown that Lys112 Lys114 cofilin crucially but differently involved in its interaction with actin phosphatidylinositol 4,5-bisphosphate. This the first example among whose inactivate their...

Cofilin, a 21kDa actin-binding protein, binds to F-actin in 1 : molar ratio of cofilin actin molecule (Nishida, E., S. Maekawa, and H. Sakai, Biochemistry, 23, 5307-5313, 1984) is capable controlling polymerization depolymerization vitro pH-sensitive manner (Yonezawa, N., E. Nishida, J. Biol. Chem., 260, 14410-14412, 1985). In this study, immunoblot analysis using monospecific antibodies against showed that ubiquitously distributed variety bovine rat organs tissues. Cofilin also present...

Cofilin is an F-actin side-binding and -depolymerizing protein with apparent molecular mass of 21 kDa. By means the end label fingerprinting method, amino acid residue on cofilin sequence cross-linked to actin by zero length cross-linker, 1-ethyl-3-(3-dimethylamino propyl)carbodiimide, was identified as Lys112 and/or Lys114. A synthetic dodecapeptide patterned around actin-cross-linking site (Trp104-Met115) inhibited binding actin. Moreover, found be a potent inhibitor polymerization. Thus,...

Cofilin, a 21-kDa actin-binding protein, has hexapeptide sequence DAIKKK which is identical to the N-terminal portion (residues 2-7) of tropomyosin. The synthetic heptapeptide, DAIKKKL, corresponding residues 122-128 cofilin, inhibited binding cofilin F-actin in dose-dependent manner. heptapeptide cosedimented with F-actin, decreased fluorescence intensity pyrene-labeled and increased rate polymerization G-actin. hexapeptides, DIKKKL DAIKKL, also affected actin polymerization, like...

This paper describes the development of a novel multi-wheel stair-climbing wheelchair. The necessity for mobility aid technology elderly and handicapped people that has "minimal invasiveness use in an historical environment" is described. With this goal mind, prototype wheelchair having function resulting from transformable wheeled four-bar linkages proposed. mechanical design, principle operation statics proposed mechanism are thoroughly illustrated. basic performance been confirmed through...

Cofilin is a 21,000-Mr actin-binding protein that widely exists in mammalian tissues. (1) A new purification procedure for porcine brain cofilin has been developed involves (NH4)2SO4 fractionation and sequential chromatographies on Toyo Pearl butyl-Toyo hydrophobic columns, hydroxyapatite, phosphocellulose Sephadex G-75 gel-filtration columns. The purified bound to F-actin increased the amount of G-actin limited extent, as previously reported [Nishida, Maekawa & Sakai (1984) Biochemistry...

The three glycoproteins of pig zona pellucida (ZPA, ZPB and ZPC) can be separated into ZPA a mixture ZPB/ZPC by gel‐filtration HPLC. We have shown previously that the neutral complex‐type N‐linked carbohydrate chains obtained from possess sperm‐binding activity. Intact ZPC cannot each other unless acidic N ‐acetyllactosamine regions their are removed endo‐β‐galactosidase digestion. endo‐β‐galactosidase−digested retains Recently, we reported N‐terminal fragment (residues 137−247) involved...

Abstract It has been proposed that mammalian sperm bind species‐specifically to carbohydrate chains of zona pellucida glycoproteins at fertilization. Although the ligand have characterized in mice and pigs, existence ligands other mammals remains unclear. In order explore bovine ligand, two vitro competition assay methods were applied. As a result, high‐mannose‐type chain, Manα1‐6(Manα1‐3)Manα1‐6(Manα1‐3)Manβ1‐4GlcNAcβ1‐4GlcNAc, which is major neutral chain egg glycoproteins, was shown...

The N‐linked oligosaccharides that were released by hydrazinolysis from glycoproteins of zonae pellucidae bovine ovarian eggs, composed neutral (23%) and acidic (77%) carbohydrate chains; almost all the chains neutralized sialidase digestion. Sugar mapping analysis pyridylaminated reverse‐phase normal‐phase HPLC 500‐MHz 1 H‐NMR spectroscopy revealed major chain is a high‐mannose‐type oligosaccharide are di‐, tri‐, tetra‐antennary, fucosylated complex‐type have N ‐acetyllactosamine repeats in...

Summary The zona pellucida (ZP) is a transparent envelope that surrounds the mammalian oocyte and mediates species-selective sperm–egg interactions. Porcine bovine ZPs consist of glycoproteins ZP2, ZP3, ZP4. In both pig heterocomplex consisting ZP3 ZP4 binds to sperm, however it not clarified whether or in complex responsible for sperm binding. Previously, we have established baculovirus-Sf9 cell expression system porcine ZP glycoproteins. A mixture recombinant (rZP3) rZP4 displayed...

The time for solubilization of the bovine zona pellucida in a hypotonic buffer containing 5% (v/v) β-mercaptoethanol and 7 mol urea l−1 increased by 10% after fertilization. Coupling with specific fluorescent thiol probe, monobromobimane (mBBr), was markedly greater ovarian eggs compared fertilized eggs, indicating that cysteine residues unfertilized are oxidized to cystines during After endo-β-galactosidase digestion remove N-acetyllactosamine repeats carbohydrate chains, three...

Abstract The zona pellucida (ZP) surrounding the mammalian oocyte is composed of three glycoprotein components (ZPA, ZPB, and ZPC). Mammalian sperm bind to carbohydrate chains a ZP in initial phase fertilization. Sperm‐ligand have been characterized mouse, cow, pig. In pigs, triantennary/tetraantennary neutral complex‐type from ZPB/ZPC mixture possess stronger sperm‐binding activity than those biantennary (Kudo et al., 1998: Eur J Biochem 252:492–499). Most these oligosaccharides...

We describe the purification of an actin regulatory protein from bovine adrenal medulla. This caused a dose-dependent decrease specific viscosity solution within 30 s its addition in Ca2+-sensitive way. Sedimentation assays and observation by electron microscopy showed that this effect was ascribable to fragmentation filaments. apparently promoted nucleation polymerization increased critical concentration for nearly 5-fold, suggesting binding barbed end The inhibitory on elongation myosin...

Short tandem repeat studies are powerful tools for parentage analysis and identification of missing persons, victims murder, mass fatalities when reference samples unavailable. The primer in the Identifiler kit failed to amplify an allele at D19S433 locus, producing a silent ("null") allele. causal mutation is base change (G>A) 32 nucleotides downstream from 3' end AAGG repeats. alleles problematical because transmitted, they can cause parent-child inconsistency that unrelated Mendelian...