An isopyoverdin from Pseudomonas putida CFML 90-44

From Pseudomonas putida CFML 90-44 an isopyoverdin was isolated. Its structure could be elucidated by chemical degradation and spectroscopic data.     

Prion protein 90-231 contains a streptavidin-binding motif

The biological function of prion protein (PrP) and the physiological relevance of its truncated subtypes and glycoforms is still enigmatic. In this paper, we adduce evidence that recombinant murine PrP fragment 90-231 (mPrP90-231) contains a biotin-mimicking sequence motif that causes binding of the bacterial protein streptavidin to mPrP90-231. As indicated by epitope mapping and proven by analysis of a deletion mutant (mPrP101-231), streptavidin binding is primarily mediated by the amino-terminus of mPrP90-231 with the core-binding sequence represented by residues 94-100. Competition with biotin significantly reduces the interaction pointing to an involvement of streptavidin's biotin-binding site (BBS). Since the BBS of streptavidin shares similarities with the active sites of proteins involved in biotin metabolism we speculate that biotin mimicry by truncated PrP-species may have an impact in vivo.     

Bat rabies in South Carolina, 1970-90.

This retrospective study examines the geographic and temporal distribution of bat species submitted for rabies testing in South Carolina (USA) from 1970 to 1990. Histories of human or animal exposures to rabid and nonrabid bats submitted during this time period were compared. Two hundred and thirty-one bats were found to be rabid from the 2,657 bats submitted over this 20 yr period. The two species most frequently submitted for testing were Lasiurus borealis with 785 specimens (30% of the total) and Nycticeius humeralis with 607 specimens (22% of the total). Lasiurus borealis also had the highest prevalence of rabies (18%) while N. humeralis had the lowest prevalence (3%). Fifty-one percent (1,259) of the bats received for testing were submitted from June through August. The majority (54%) of rabid bats were received from August through October.     

Hsp90 & Co. - a holding for folding.

Hsp90 is an abundant molecular chaperone that is involved in the folding of a defined set of signalling molecules including steroid-hormone receptors and kinases. Recent in vitro experiments suggest that Hsp90 contains two different binding sites for non-native proteins, which allow it to combine the properties of a promiscuous chaperone with those of a dedicated folding-helper protein. Significant progress has been made in analysing co-chaperones, which form defined, substrate-dependent complexes with Hsp90 in vivo. Structural studies have identified the ATP-binding site in the N-terminal domain of Hsp90, which can be blocked by high-affinity inhibitors. Although a detailed understanding of the mechanism of Hsp90 action is still lacking, recent advances suggest that the protein is the centre of a dynamic, multifunctional and multicomponent chaperone machinery that extends the limits of protein folding in the cell.     

Mn-peroxidase from Bjerkandera adusta 90-41. Purification and substrate specificity

Mn-peroxidase has been purified to homogeneity from culture liquid of white-rot fungus Bjerkandera adusta 90-41 grown on medium containing lignosulfonates. According to the data on SDS-PAGE and isoelectrofocusing, the molecular mass of the enzyme is 43 kD and the isoelectric point is 3.5. The pH-optimum in the reaction of MnSO4 oxidation is 4.5. The substrate specificity of the enzyme has been studied. In contrast to previously known Mn-peroxidases from B. adusta, the isolated enzyme has no activity with veratryl alcohol. The enzyme can oxidize ammonium 2, 2-azino-bis(ethyl-6-benzothiazoline sulfonate) (ABTS), o-phenylenediamine, and phenol red in the absence of Mn2+. Oxidation of ABTS and o-phenylenediamine is stimulated by Mn2+, whereas in the reaction of oxidation of phenol red Mn2+ acts as an inhibitor. Some aromatic substrates, such as pyrocatechol and guaiacol, are oxidized only in the presence of Mn2+.     

Bacterial infection and tissue-specific Hsp72, -73 and -90 expression in western painted turtles

Heat shock proteins (Hsps) are molecular chaperones that assist intracellular folding, assembly and translocation of proteins in prokaryotic and eukaryotic cells. A variety of stresses including hyperthermia, radiation, heavy metals, ischemia, anoxia and reoxygenation have been shown to increase the expression of Hsps. Likewise, bacterial infection represents a stress for the host cell. In this study, expression of the constitutive (Hsp73) and inducible (Hsp72) isoforms of Hsp70 and Hsp90 was monitored in brain, heart, liver and skeletal muscle from the western painted turtle Chrysemys picta bellii diagnosed with Septicemic Cutaneous Ulcerative Dermatitis (SCUD). This disease is caused by a gram-negative bacterium probably belonging to the Citrobacter spp. The expression of Hsp73 increased 1.8-fold in brain and liver, 2.2-fold in heart but did not change in skeletal muscle; Hsp72 expression increased 5.5-fold in brain and 3-fold in liver but did not change in heart or skeletal muscle; Hsp90 expression increased 9-fold in brain, 2.7-fold in heart and 2.4-fold in skeletal muscle but did not change in liver. These results suggest a tissue-specific Hsp response during bacterial infection and a role for Hsps in immunopathological events in reptiles.     

Haematological studies on90Sr-90Y-toxicity

Summary Erythropoietic changes were observed, measured by⁵⁹Fe-uptake into red blood cells, and on radioiron turnover from blood plasma, at different time intervals (2–64 days) after treating adult female mice with varying activities of⁹⁰Sr-⁹⁰Y. Activities of 2.5 or 5.0 µCi radiostrontium per animal lead to a depression at time intervals of two and four days, at longer periods there was an overshoot. With activities of 0.5 or 1.0 µCi radiostrontium disturbances in the radioiron uptake are still observed, although these effects are not as pronounced as in experiments with higher burdens. In comparison with results obtained in experiments in which the plasma⁵⁹Fe-turnover was applied, even with an activity of 5 µCi radiostrontium per mouse, no deviation as against the untreated controls was detected.Dedicated to Prof. Dr. Hermann Muth, Homburg/Saar, on the occasion of his 65th birthday     

Asthma deaths in Cardiff 1963-74: 90 deaths outside hospital.

In a detailed study of factors associated with death from bronchial asthma outside hospital 90 patients were investigated. The fatal attack was typically short and was most likely to occur in patients with a long history. Deaths often occurred before effective medical help was obtainable, but occasionally the patient or the doctor underestimated the severity of the attack. Patients especially at risk were those recently discharged from hospital after a previous attack. These deaths might be prevented by better patient education, a self-admission service for selected asthmatics, and by doctors using objective measurements of severity of asthma for the control of treatment. The underuse of corticosteroids is an important factor associated with death.     

Solution structure of Syrian hamster prion protein rPrP(90-231)

NMR has been used to refine the structure of Syrian hamster (SHa) prion protein rPrP(90-231), which is commensurate with the infectious protease-resistant core of the scrapie prion protein PrPSc. The structure of rPrP(90-231), refolded to resemble the normal cellular isoform PrPC spectroscopically and immunologically, has been studied using multidimensional NMR; initial results were published [James et al. (1997) Proc. Natl. Acad. Sci. U.S.A. 94, 10086-10091]. We now report refinement with better definition revealing important structural and dynamic features which can be related to biological observations pertinent to prion diseases. Structure refinement was based on 2778 unambiguously assigned nuclear Overhauser effect (NOE) connectivities, 297 ambiguous NOE restraints, and 63 scalar coupling constants (3JHNHa). The structure is represented by an ensemble of 25 best-scoring structures from 100 structures calculated using ARIA/X-PLOR and further refined with restrained molecular dynamics using the AMBER 4.1 force field with an explicit shell of water molecules. The rPrP(90-231) structure features a core domain (residues 125-228), with a backbone atomic root-mean-square deviation (RMSD) of 0.67 A, consisting of three alpha-helices (residues 144-154, 172-193, and 200-227) and two short antiparallel beta-strands (residues 129-131 and 161-163). The N-terminus (residues 90-119) is largely unstructured despite some sparse and weak medium-range NOEs implying the existence of bends or turns. The transition region between the core domain and flexible N-terminus, i.e., residues 113-128, consists of hydrophobic residues or glycines and does not adopt any regular secondary structure in aqueous solution. There are about 30 medium- and long-range NOEs within this hydrophobic cluster, so it clearly manifests structure. Multiple discrete conformations are evident, implying the possible existence of one or more metastable states, which may feature in conversion of PrPC to PrPSc. To obtain a more comprehensive picture of rPrP(90-231), dynamics have been studied using amide hydrogen-deuterium exchange and 15N NMR relaxation times (T1 and T2) and 15N{1H} NOE measurements. Comparison of the structure with previous reports suggests sequence-dependent features that may be reflected in a species barrier to prion disease transmission.     

Protease and alpha-amylase synthesis by washed cells of Aspergillus oryzae 251-90

Regularities of protease and alpha-amylase production by washed cells of Aspergillus oryzae 251-90 were being studied. The results obtained enabled us to assume a constitutive character of the both enzymes synthesis by the given producer. Sources of carbon, nitrogen and sulphur take part in regulation of the protease production, whereas in the case of the alpha-amylase synthesis only carbon sources that are important. Elimination of phosphorus from the medium affects the synthesis of both enzymes. Celatin stimulates the production of the two enzymes, being a supplier of amino acids.