Purification and characterization of a vitamin D3 25-hydroxylase from pig liver microsomes.

A cytochrome P-450 which catalyses 25-hydroxylation of vitamin D3 has been purified to apparent homogeneity from pig liver microsomes. The specific content of cytochrome P-450 was 12 nmol.mg of protein-1, and the preparation showed a single band with an apparent M(r) of 50,500 upon SDS/PAGE.

Variant forms of galanin isolated from porcine brain.

In a peptide concentrate, prepared from acid extracts of porcine brain, several galanin-like immunoreactive peptides were detected and two of these were purified. Characterization of the peptides by sequence analysis, mass spectrometry, and capillary zone electrophoresis identified them as a N-terminally nine residue elongated form of galanin, preprogalanin(24-61) amide, and as an N-terminally four residue truncated form of galanin corresponding to preprogalanin(37-61) amide. The former finding suggests that the removal of the signal peptide in preprogalanin occurs by enzymatic cleavage between glycine-23 and leucine-24.

Results of a prospective randomized trial for treatment of severely brain-injured patients with hyperbaric oxygen.

The authors enrolled 168 patients with closed-head trauma into a prospective trial to evaluate the effect of hyperbaric oxygen in the treatment of brain injury. Patients were included if they had a total Glasgow Coma Scale (GCS) score of 9 or less for at least 6 hours. After the GCS score was established and consent obtained, the patient was randomly assigned, stratified by GCS score and age, to either a treatment or a control group.

A synthetic peptide encompassing the binding site of the second zinc atom (the 'structural' zinc) of alcohol dehydrogenase.

A 23-residue peptide was synthesized that incorporates the loop which binds the structural zinc atom of mammalian alcohol dehydrogenases and contributes, in part, to subunit interactions in the native enzyme. Neither the amino acid composition nor the sequence of the peptide resemble those of zinc fingers. The reduced peptide stoichiometrically binds zinc or cobalt to form stable complexes with a dissociation constant of the peptide/CO2+ complex of 2.

Amino acid sequence of PR-39. Isolation from pig intestine of a new member of the family of proline-arginine-rich antibacterial peptides.

We recently isolated from pig intestine and characterized a 31-residue antibacterial peptide named cecropin-P1 with activity against Escherichia coli and several other Gram-negative bacteria. The isolation involved a number of batch-wise steps followed by several chromatography steps. The continued investigation of these antibacterial peptides has now yielded another antibacterial peptide with high activity against both E.

Subependymoma of the cervical spinal cord. A case report of long-term survival.

Subependymomas of the cervical spinal cord are relatively rare tumors. Thirteen cases have been reported in the literature with the longest reported follow-up and survival being six years. This paper describes a woman who survived for 14 years in reasonably good health after her diagnosis of cervical spinal cord intramedullary subependymoma.

Characterization of mitochondrial cytochromes P-450 from pig kidney and liver catalysing 26-hydroxylation of 25-hydroxyvitamin D3 and C27 steroids.

The properties of cytochrome P-450 from pig kidney mitochondria, catalysing 26-hydroxylation of 25-hydroxyvitamin D3 and C27 steroids [Postlind & Wikvall (1989) Biochem. Biophys. Res.

Direct analysis of peptides and amino acids from capillary electrophoresis.

Capillary zone electrophoresis of peptides on a preparative scale and direct analysis of both amino acid sequences and compositions are demonstrated. Using native and synthetic peptides prepared by repetitive runs and appropriate collections, sufficient material for direct sequence analysis is obtained after just a few runs, while single runs suffice for total compositions. Electrophoretic direct amino acid analysis without derivatization is also possible and gives especially high sensitivity for aromatic residues (less than 200 fmols) plus convenient distinction of amidated versus free C-terminal residues after carboxypeptidase digestions.

Leukotriene A4 hydrolase in the human B-lymphocytic cell line Raji: indications of catalytically divergent forms of the enzyme.

Leukotriene A4 hydrolase was purified 1400-fold, with an approximate yield of 25%, to apparent homogeneity from the human B-lymphocytic cell line Raji. The purification included ammonium sulfate precipitations followed by anion exchange, hydrophobic interaction, and molecular exclusion fast protein liquid chromatography. Kinetic properties at 2 degrees C varied between different enzyme preparations.

Characterization of a tartrate-resistant acid phosphatase (ATPase) from rat bone: hydrodynamic properties and N-terminal amino acid sequence.

Certain physicochemical properties of rat bone tartrate-resistant acid ATPase (TrATPase), including the size and shape of the enzyme, potential subunit composition, and detergent binding, have been elucidated. SDS-polyacrylamide gel electrophoresis in combination with immunoblot analysis showed that the bone TrATPase has a molecular weight of 33,000 D and is composed of disulfide-linked polypeptides of 20,000 and 16,000 D. The enzyme contains 1.

Characterization of human deoxycytidine kinase. Correlation with cDNA sequences.

Existing data on the structure of human deoxycytidine kinase (dCK) diverge. A monomeric 60 kDa form has been isolated and the cloning of a cDNA coding for 626 amino acids corresponding to a 71 kDa protein has been reported. However, pure dCK isolated from leukemic spleen is a dimer of 30 kDa subunits.

Analysis of the V antigen lcrGVH-yopBD operon of Yersinia pseudotuberculosis: evidence for a regulatory role of LcrH and LcrV.

Virulent Yersinia species possess a common plasmid that encodes essential virulence determinants (Yops) which are regulated by the extracellular stimuli Ca2+ and temperature. The V antigen operon was recently shown to be involved in the Ca2(+)-regulated negative pathway (A. Forsberg and H.

Eukaryotic glucose-6-phosphate dehydrogenases: structural screening of related proteins.

Rapid assessment of structural relationships between yeast glucose-6-phosphate dehydrogenases and other eukaryotic types of this enzyme is described. Separation and size estimation of large fragments by sodium dodecylsulfate/polyacrylamide gel electrophoresis, electroblotting onto disks, and sequencer analysis provide data that permit alignment of the segments thus characterized with the related proteins, and utilize existing structural knowledge to assess new enzyme structures. Affinity labeling allows further correlations.

A fragment of triosephosphate isomerase competes with the vasoactive intestinal polypeptide (VIP) for binding to the VIP receptor.

A 28-residue N-terminal fragment of triosephosphate isomerase, TIM(1-28), has been purified from porcine upper intestine. It competes with VIP for binding to the VIP receptor on rat liver plasma membranes with an IC50 value of 2.8 mM, about 1000 times higher than that for VIP binding to the membranes.

Characterization of pig kidney microsomal cytochrome P-450 catalysing 25-hydroxylation of vitamin D3 and C27 steroids.

The cytochrome P-450 enzyme which catalyses 25-hydroxylation of vitamin D3 (cytochrome P-450(25] from pig kidney microsomes [Postlind & Wikvall (1988) Biochem. J. 253, 549-552] has been further purified.

Antibodies with specificities against a dispase-produced 15-kilodalton hexon fragment neutralize adenovirus type 2 infectivity.

During the entrance of adenovirus type 2 into cells, it has been suggested that the virion undergoes a conformational change. In this investigation, we have further characterized the hypothetical conformational change, which the structural protein hexon undergoes in response to low pH. From pH 5.

Halo immobilization and surgical fusion: relative indications and effectiveness in the treatment of 140 cervical spine injuries.

In the management of cervical spine injuries, it is not always clear when to use halo immobilization alone, surgical fusion alone, or a combination of the two. To investigate the relative effectiveness of each of these approaches, we reviewed the medical records of 140 patients with cervical spine injuries treated with either halo immobilization or surgical fusion, or both. Seventy (50%) of the patients were neurologically intact on admission (two of these were paraplegic from previous injuries).

Electroblotting of polypeptides onto glass fiber filters for direct sequence analysis after sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

The technique of electroblotting polypeptides onto Polybrene-treated glass fiber filter discs after protein detection with potassium chloride is evaluated further with different proteins in separate applications. The number of proteins analyzed with this method is now more than double that previously reported. Reproducible results in good yield are obtained.

A new member of a secretory protein gene family in the dipteran Chironomus tentans has a variant repeat structure.

We describe the structure of a gene expressed in the salivary gland cells of the dipteran Chironomus tentans and show that it encodes 1 of the approximately 15 secretory proteins exported by the gland cells. This sp115,140 gene consists of approximately 65 copies of a 42-bp sequence in a central uninterrupted core block, surrounded by short nonrepetitive regions. The repeats within the gene are highly similar to each other, but divergent repeats are present in a pattern which suggests that the repeat structure has been remodeled during evolution.

The cytotoxic protein YopE of Yersinia obstructs the primary host defence.

It has previously been shown that the plasmid-encoded YopE protein of Yersinia pseudotuberculosis is a virulence determinant. In this study, HeLa cells, macrophages and mice were used as different model systems to determine the actual role of YopE in the virulence process. The YopE protein mediates a cytotoxic response on a confluent layer of HeLa cells.

Rat and porcine galanin are equipotent in inhibiting insulin responses to glucose in the anesthetized rat.

The structure of rat galanin, recently elucidated using recombinant DNA technology, differs from porcine galanin by three amino acid residue substitutions at positions located in the C-terminal region. A synthetic replicate of the proposed structure of rat galanin was prepared and its potency to inhibit insulin responses to glucose in anesthetized rats was compared with that of porcine galanin. Within experimental error, the dose-response curves of porcine and rat galanin to inhibit glucose-stimulated rat insulin responses were indistinguishable.

Identification of Gly-Pro-Glu (GPE), the aminoterminal tripeptide of insulin-like growth factor 1 which is truncated in brain, as a novel neuroactive peptide.

A truncated form of IGF-1 which lacks the aminoterminal tripeptide Gly-Pro-Glu (GPE) is found in human brain. It was proposed that GPE may result from neural specific processing and also have a function within the CNS. GPE was synthesized and shown to inhibit glutamate binding to the N-methyl-D-aspartate (NMDA) receptor.