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Biomedical subjects

P Crine

Publications and source records attributed to P Crine.

118 records · Page 7Linked to original sources
PubMed1977

In vitro biosynthesis of beta-endorphin in pituitary glands.

beta-Endorphin is a 31 amino acid polypeptide isolated from the pituitary gland of different species of animals. It has strong morphine-like activity. It is formed of amino acid residues 61-91 of beta-lipotropin. Speculation has arisen whether it is biosynthesized in situ or transformed after secretion of beta-lipotropin. The present in vitro studies show that it is found as beta-endorphin in bovine pituitary slices incubated with radioactive amino acid precursor [35S]methionine. Chemical characterization and microsequencing of the newly biosynthesized material proves its identity with isolated unlabeled beta-endorphin and shows that it has a methionine residue at its fifth position, as expected.

Amino Acids↗
PubMed1976

A study of DNA spontaneous degradation.

The intact apurinic site half-life in DNA molecules, at 37 degrees C and pH 7.21, in a medium approximating the intracellular ionic composition, was found to be between 288 and 335 h. The spontaneous degradation of 3H-labelled T7 phage DNA, at 37 degrees C and pH 7.21, in the same medium, was followed by sedimentation analysis on sucrose gradients after denaturation with NaOH; it was found to be equivalent to 0.009 break per h per DNA strand. The number of intact apurinic sites found in this incubated T7 phage DNA was much lower than the theoretical value calculated on the assumption that depurination was the only cause of DNA degradation. We concluded that depurination was a minor cause of DNA degradation in our experimental conditions.

Chemical Phenomena↗
PubMed1974

In vitro repair of apurinic sites in DNA.

Apurinic sites disappear from DNA during an incubation of the DNA with the Escherichia coli endonuclease specific for apurinic sites, DNA polymerase I (EC 2.7.7.7.), and T4 ligase (EC 6.5.1.1). Omission of any one of these three enzymes and, in particular, omission of the endonuclease specific for apurinic sites prevents this in vitro repair.

Binding Sites↗
PubMedDate not supplied

Characterization of sulfated forms of the pro-opiomelanocortin amino-terminal glycopeptide in rat intermediate lobe cells.

Explants of rat neurointermediate lobes were incubated in the presence of radioactive amino acids, sugars or sulfate and the labeled proteins were separated by two-dimensional gel electrophoresis. A double series of acidic peptides (Mr = 16,000-21,500) were identified as variant forms of the amino-terminal glycopeptide of pro-opiomelanocortin (N-POMC). The series of peptides with the higher molecular weights (Mr = 18,000-21,500) contain a tryptic fragment (tentatively identified as the tryptic peptide of the "joining peptide": sequence 77 to 93 of rat POMC) which is absent from the forms of the lower molecular weight series (Mr = 16,000 to 18,000). Pulse-chase studies further showed that the high molecular weight forms of N-POMC could be post-translationally cleaved albeit slowly into the species of Mr = 16,000-18,000 which constitute, at least in part, the final maturation products of the N-terminal region of the precursor molecule. All the variant forms of the N-POMC glycopeptide could be labeled with [35S]sulfate. Our results strongly suggest that most of the sulfate groups are attached to N-linked oligosaccharide side chains of N-POMC. We therefore propose that one of the final maturation products of the N-terminal portion of POMC in rat intermediate lobes is a sulfated glycopeptide (Mr = 16,000-18,000) composed of the 1-74 sequence of rat POMC.

Amino Acid Sequence↗
PubMedDate not supplied

Complete sequence of rabbit kidney aminopeptidase N and mRNA localization in rabbit kidney by in situ hybridization.

We have isolated and sequenced a full-length cDNA for rabbit kidney aminopeptidase N (APN). The 3-kilobase cDNA contains 12 nucleotides of the 5' noncoding region, a 2898 nucleotide long open reading frame, and 113 nucleotides of the 3' untranslated region. The open reading frame encodes a type II membrane protein of 966 amino acid residues composed of a 10 residue NH2-terminal cytosolic domain, a 23 residue transmembrane domain, and a large 933 residue ectodomain that contains the active site. Rabbit APN has eight potential N-glycosylation sites and seven cysteine residues, one of which is located in the transmembrane domain. Computer analysis showed that the enzymes from human, rat, and rabbit were highly conserved, except for the stalk region immediately downstream from the transmembrane domain. Using in situ hybridization techniques we showed that in rabbit kidney, APN mRNA is present in proximal tubules but not in glomeruli, which corresponds to the localization of the protein observed by immunohistochemistry. Taken together, our results strongly suggest that the expression of APN in kidney is modulated at mRNA levels and not at translational and (or) posttranslational levels.

Amino Acid Sequence↗
PubMedDate not supplied

New perspectives in neuropeptides. Biosynthesis from large precursors. The pro-opio-melanocortin model in the pars intermedia.

Although more and more is known about the nature of neuropeptides, there are two basic questions which need to be looked at in order for the neurophysiologists and the neuropharmacologists to have all the tools necessary for their studies. The first question is the site of biosynthesis of neuropeptides and the second is their biosynthetic pathway. We believe and propose that all neuropeptides will be formed from a larger molecular weight precursor similar to beta-LPH for beta-endorphin. All models of peptide hormones (pro-insulin, pro-PTH, pro-glucagon, beta-lipotropin etc.,) show the same characteristic preferential site of cleavage made of a double basic residue. This is also the type of cleavage recently described by Nakanishi et al. (1979). Using the rat pars intermedia, we showed that this tissue preferentially makes alpha-MSH and beta-endorphin from a large precursor, while beta-LPH and ACTH are short lived intermediates. All products of maturation were identified by chemical analyses leaving no doubt about their nature and identity. We strongly believe that the brain will biosynthesize beta-endorphin and possibly alpha-MSH in similar fashion. We also predict that our results will apply to the other neuropeptides which wil be formed from large precursors. These studies could be very useful in the interpretation of the intricate relationship between neuropeptides and behavioral effects.

Adrenocorticotropic Hormone↗