A low-cost gradient programmer.
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Biomedical subjects
Publications and source records attributed to R V Lewis.
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A previously unknown peptide, betaH-Leu5-endorphin, has been reported in the dialysates of schizophrenic patients. Accordingly, hemofiltrates from two schizophrenic and two control patients were examined for the presence of betaH-Leu5-endorphin. The opioid peptides were detected by a radioreceptor assay after separation and identification by gel filtration and high-performance liquid chromatography. With a detection limit of 30 pmole/L of hemofiltrate, no betaH-Leu5-endorphin or Met5-endorphin was found in controls or in patients. Whatever the possible involvement of endorphins in schizophrenic behavior, they are not present at detectable levels in the hemofiltrates of two well-characterized schizophrenic patients, thereby casting doubt on a general relationship of Leu-endorphin and schizophrenia.
Bovine adrenal chromaffin granules have been shown to contain, in addition to Met-enkephalin and Leu-enkephalin, at least three small peptides with opiate receptor activity. One of these adrenal peptides has been purified to homogeneity and its sequence was shown to be Met-enkephalin-[Arg6,Phe&]. This heptapeptide was also found in beef striatal extracts in amounts comparable to those of Leu-enkephalin.
Pro-opiocortin was purified from camel pituitaries by procedures including high-performance liquid chromatography. The precursor relationship of the pure protein to the opioid peptides and to corticotropin was confirmed. Partial chemical analysis consisting of amino acid analysis and tryptic peptide mapping was carried out with the aid of sensitive fluorescence detection.
Chromatographic procedures have been developed for resolving all of the known enkephalins and endorphins on a single column. The effect of eluant pH on the retention times and separation of the enkephalins and beta-endorphin was determined. By combining these separations with a sensitive radioreceptor assay it is possible to assay all of the opioid peptides in the pituitary gland or in various regions of the brain from individual small laboratory animals.
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Striatal extracts of guinea pigs, rats, and cattle were found to contain two large proteins (greater than 40,000 and greater than 100,000 daltons) that on treatment with trypsin yield opioid peptides differing chromatographically from the opioid nonapeptide generated by trypsin digestion of endorphins, beta-lipotropin, or pro-opiocortin. Furthermore, the large opioid proteins found in the pituitary do not appear to be present in the striatum. These and other findings indicate that the striatal enkephalins are produced via a pathway from the one deduced from studies on the pituitary.
Two novel heterobifunctional cross-linking reagents, which can be used to attach photoactivatable nitroaryl azides to primary amino groups of proteins, have been synthesized. The two compounds, N-5-azido-2-nitrobenzoyloxy-succinimide and ethyl N-5-azido-2-nitrobenzoylaminoacet-imidate-HCl, as well as ethyl 4-azidobenzimidate-HCl have been attached to lysine residues of cobra venom phospholipase A2 without a loss in enzymatic activity. Subsequent illumination of the modified forms of the enzyme at appropriate wavelengths under conditions in which the native enzyme exists in an aggregated state led to the formation of covalently linked dimers and large aggregates which could be separated by electrophoresis on polyacrylamide gels in the presence of sodium dodecyl sulfate.
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