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P E Butler

Publications and source records attributed to P E Butler.

54 records · Page 3Linked to original sources
PubMed1988

A latent proteinase in mouse kidney membranes. Characterization and relationship to meprin.

Inbred mice can be phenotypically divided into two groups: those that contain high levels of a kidney metallo-endopeptidase activity (meprin-a) and those with low meprin-a activity. In studies to investigate the molecular basis for the heterogeneity in the expression of this proteinase activity, we found a latent metallo-proteinase activity associated with kidney membranes of C3H/HeJ mice, a low activity strain. The latent proteinase was activated by treatment of kidney brush border membranes with trypsin and was purified from solubilized C3H kidney membranes. Purified preparations of the C3H latent proteinase (referred to as meprin-b) contained three major proteins of subunit molecular weights 90,000, 140,000, and 160,000. In the absence of reducing agents, four 90,000-Da subunits are covalently linked by S-S bridges. The two higher molecular mass proteins are not covalently linked to each other or to the 90,000-Da subunits. However, cross-linking and affinity chromatography studies indicated that the proteins in the meprin-b preparation were tightly associated. By contrast, purified meprin-a contains only 85,000-Da subunit proteins linked by S-S bridges to form a tetramer. Endoglycosidase F treatment decreased the mass of the 90,000-Da meprin-b subunit and the 85,000-Da meprin-a subunit to polypeptides of 65,000-70,000 Da. The 90,000- and 85,000-Da subunits are immunologically similar, in that polyclonal antibodies prepared against one of the subunits cross-react with the other. The substrate specificities and inhibitor profiles of purified preparations of meprin-a and meprin-b are also similar. These data are consistent with the proposition that meprin-b is a polymorphic form of meprin-a that is incompletely processed in vivo.

Amino Acids, Sulfur↗
PubMed1987

Characterization of meprin, a membrane-bound metalloendopeptidase from mouse kidney.

Meprin is an intrinsic protein of the brush border, a specialized plasma membrane, of the mouse kidney. It is a metalloendopeptidase that contains 1 mol of zinc and 3 mol of calcium per mol of the 85,000-Mr subunit. The enzyme is isolated, and active, as a tetramer. The behaviour of the enzyme on SDS/polyacrylamide gels in the presence and absence of beta-mercaptoethanol indicates that the subunits are of the same Mr (approx. 85,000) and held together by intersubunit S--S bridges. Eight S-carboxymethyl-L-cysteine residues were detected after reduction of the enzyme with beta-mercaptoethanol and carboxymethylation with iodoacetate. The enzyme is a glycoprotein and contains approx. 18% carbohydrate. Most of the carbohydrate is removed by endoglycosidase F, indicating that the sugar residues are N-linked. The isoelectric point of the enzyme is between pH 4 and 5, and the purified protein yields a pattern of evenly spaced bands in this range on isoelectric focusing. The peptide-bond specificity of the enzyme has been determined by using the oxidized B-chain of insulin as substrate. In all, 15 peptide degradation products were separated by h.p.l.c. and analysed for their amino acid content and N-terminal amino acid residue. The prevalent peptide-bond cleavages were between Gly20 and Glu21, Phe24 and Phe25 and between Phe25 and Tyr26. Other sites of cleavage were Leu6-Cysteic acid7, Ala14-Leu15, His10-Leu11, Leu17-Val18, Gly8-Ser9, Leu15-Tyr16, His5-Leu6. These results indicate that meprin has a preference for peptide bonds that are flanked by hydrophobic or neutral amino acid residues, but hydrolysis is not limited to these bonds. The ability of meprin to hydrolyse peptide bonds between small neutral and negatively charged amino acid residues distinguishes it from several other metalloendopeptidases.

Amino Acids↗
PubMed1986

Metalloendopeptidases of the mouse kidney brush border: meprin and endopeptidase-24.11.

Two metalloendopeptidases, meprin and endopeptidase-24.11 ("24.11"), were isolated from mouse kidney membranes, and their structural and catalytic properties were investigated. The enzymes both cross-react with antibodies prepared in rabbits against purified preparations of meprin; thus they share some immunologic determinants. Meprin and 24.11 have similar subunit molecular weights of 85 000 and 90 000, respectively, as demonstrated after sodium dodecyl sulfate polyacrylamide gel electrophoresis in the presence of 2-mercaptoethanol. However, under non-reducing conditions, meprin migrates as an oligomer while 24.11 remains monomeric. This and other data indicate that meprin subunits are linked by disulfide bridges, whereas endopeptidase-24.11 subunits are not covalently linked. Both endopeptidases hydrolyze insulin B chain and are totally inhibited by EDTA and o-phenanthroline. The activity of 24.11 against insulin B chain was totally inhibited by low concentrations of phosphoramidon (less than 2 nM), whereas meprin was not inhibited by concentrations of this inhibitor as high as 20 microM. Large proteins are not substrates for endopeptidase-24.11, while meprin degrades proteins such as azocasein rapidly (apparent Km = 0.65 mg/ml). Meprin appears to require an extended polypeptide chain in substrates while 24.11 prefers smaller peptides as substrates. Both endopeptidases have a preference for peptide bonds that contain hydrophobic amino acids. With the octapeptide angiotensin II as substrate, both enzymes hydrolyze the central Tyr-Ile bond; 24.11 also cleaves at Arg-Val and Ile-His. The two endopeptidases show many similarities immunologically, structurally and catalytically, however, they display distinct characteristics which may be physiologically important.

Angiotensin II↗
PubMed1985

The turnover of skeletal muscle glycogen phosphorylase studied using the cofactor, pyridoxal phosphate, as a specific label.

The turnover of glycogen phosphorylase has been measured using the cofactor, pyridoxal phosphate, as a label specific for this enzyme in skeletal muscle. Radiolabelled pyridoxine administered in vivo is incorporated into a protein-bound fraction in skeletal muscle, shown by several criteria to be equivalent to glycogen phosphorylase. This pool of radiolabel disappears slowly with a half-life of 11.9 days, taken to be a good estimate of the intracellular half-life of the enzyme. The use of the cofactor in this fashion minimises overestimation of half-life that results from reincorporation of the label. Further, premature dissociation of the cofactor from native enzyme, which would lead to underestimation of half-life, is unlikely. At the level of sensitivity given by this method there was little evidence for the appearance of pyridoxal phosphate-labelled degradation intermediates of the enzyme.

Animals↗
PubMed1985

Accelerated degradation of glycogen phosphorylase in denervated and dystrophic mouse skeletal muscle.

Pyridoxal phosphate, the cofactor of glycogen phosphorylase, fulfils the criteria needed of a turnover label for this enzyme. The decay of protein-bound label following administration of [3H]pyridoxine is a good index of the rate of degradation of the enzyme in vivo. This method has been applied to the study of catabolism of the enzyme in normal, denervated and dystrophic mouse skeletal muscle. In both of the pathological conditions the enzyme is degraded more rapidly than normal.

Animals↗
PubMed1985

Nurses' uniform: an investigation of mobility.

An investigation of the mobility of nurses under three clothing conditions is reported. The need for such a study has arisen as a result of the concern over a possible mismatch between mobility and patient handling requirements. Thirty-seven nurses participated under two of the clothing conditions ('National' dress uniform, Trouser/tunic combination). In addition, ten of these nurses volunteered to provide control data by being measured in a leotard or a swimming costume. Eleven static and sixteen dynamic anthropometric measures were considered. Each nurse was asked to complete a short questionnaire, relating to her subjective attitudes to the uniform and to her own physical state at the time of measurement. Whilst both uniforms imposed restrictions on the shoulder girdle and trunk of up to 10%, the area of greatest concern was the mobility of the hip joint. Hip flexion was reduced by 26% in the dress uniform. The implications of these findings for patient handling procedures are discussed, as are those of the relationship between the environment and the material. Uniform and the nursing image is also considered.

Adult↗
PubMed1984

Purification of glycogen phosphorylase from small quantities of mouse skeletal muscle.

A new approach to the purification of skeletal muscle glycogen phosphorylase is described. The purification scheme is particularly suited to preparation of the enzyme from small amounts of tissue. A combination of dye-ligand chromatography and hydrophobic chromatography yields homogenous enzyme with good recoveries. The purification is rapid and may be completed in a working day.

Acrylic Resins↗
PubMed1981

The inactivation of streptomyces-derived proteinase inhibitors by mammalian tissue preparations.

The proteinase inhibitors leupeptin and chymostatin are inactivated by preparations of perfused mouse liver. The inhibitors are degraded maximally at neutral or alkaline pH values. The inactivating enzymes are inhibited by Dip-F and pms-F but not by EDTA, 1,10 phenanthroline or iodoacetic acid. The significance of these results is discussed in terms of the potential of the inhibitors as antiproteolytic drugs.

Animals↗
PubMedDate not supplied

Prolonged general anesthesia for experimental craniofacial surgery in fetal swine.

A protocol utilizing high preoperative doses of altrenogest (Regu-Mate) and a "balanced" general anesthesia regimen consisting of isoflurane at subanesthetic doses supplemented with intravenous doses of sodium thiopental was developed to prevent preterm labor, minimize intracranial fetal cerebral edema, and decrease postpartum mortality of fetal swine after undergoing complex in utero craniofacial procedures. A total of 20 fetal piglets at 75% gestation were exposed to prolonged (> 3 h) anesthesia conditions of which 7 piglets were randomly selected to undergo experimental craniofacial procedures consisting of periosteal stripping of frontal and parietal bone segments with/without extensive coronal suture fusion procedures. Neither sows nor piglets were lost to anesthetic complications during the initial laparotomy or subsequent cesarean delivery. None of the sows experienced uterine sepsis or underwent preterm labor. The overall survival rate for all piglets exposed to prolonged anesthesia conditions was 95% at 4 weeks and 45% at 11 weeks after surgery. The experimental group's survival was 85.7% at 4 weeks and 28.5% at 11 weeks after surgery.

Anesthesia, General↗
PubMedDate not supplied

Prolonged survival in fetal rabbit surgery.

Timing and outcome of antenatal surgical intervention is being explored using fetal animal models. Models that are currently used range from larger animals with fewer offspring and higher cost to smaller animals with larger litters and lower cost. The rabbit is an ideal "small" animal model for experimentation in the third trimester, with a large litter, short gestation and a relatively large fetus. This paper reports methods by which prolonged survival (greater than 110 days) may be achieved in as many as 60% of operated fetuses following complex fetal surgery in the rabbit.

Animals↗