Alternative mRNA splicing generates multiple forms of peptidyl-glycine alpha-amidating monooxygenase in rat atrium.

Stoffers, Doris A.; Green, Cristina Barthel-Rosa; Eipper, Betty A.

doi:10.1073/pnas.86.2.735

Cited by 108 publications

(67 citation statements)

References 29 publications

(29 reference statements)

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“…Whereas in the stomach most of the gastrin cells were immunoreactive for PAL and only some were positive for PHM, in the colon only immunoreactivity for PHM was found (Martinez et al, 1993b). Tissue specific expression of alternate mRNA splice forms and post-translational proteolysis and secretion of PAM proteins have been extensively studied (Oyarce and Eipper, 1993;Stoffers et al, 1989) and could explain some of the differences found. In addition, possible differences in sensitivity between the antibodies could also explain this discrepancy.…”

Section: Discussionmentioning

confidence: 99%

Distribution of peptidyl-glycine alpha-amidating mono-oxygenase (PAM) enzymes in normal human lung and in lung epithelial tumors.

Saldise¹,

Martı́nez²,

Montuenga³

et al. 1996

J Histochem Cytochem.

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C-terminal alpha-amidation is a post-translational modification necessary for the biological activity of many regulatory peptides produced in the respiratory tract. This modification is a two-step process catalyzed by two separate enzyme activities, both derived from the peptidyl-glycine alpha-amidating mono-oxygenase (PAM) precursor. The distribution of these two enzymes, peptidyl-glycine alpha-hydroxylating monoxygenase (PHM) and peptidyl-alpha-hydroxyglycine a amidating lyase (PAL), was studied in the normal lung and in lung tumors using immunocytochemical methods and in situ hybridization. In normal lung the enzymes were located in some cells of the airway epithelium and glands, the endothelium of blood vessels, some chondrocytes of the bronchial cartilage, the alveolar macrophages, smooth muscle cells, neurons of the intrinsic ganglia, and in myelinated nerves. A total of 24 lung tumors of seven different histological types were studied. All cases contained PAM-immunoreactive cells with various patterns of distribution. All immunoreactive cells were positive for the PHM antiserum but only some of them for the PAL antiserum. The distribution of PAM co-localizes with some other previously described amidated peptides, suggesting that amidation is an important physiological process taking place in the normal and malignant human lung tissue.

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Section: Discussionmentioning

confidence: 99%

Distribution of peptidyl-glycine alpha-amidating mono-oxygenase (PAM) enzymes in normal human lung and in lung epithelial tumors.

Saldise¹,

Martı́nez²,

Montuenga³

et al. 1996

J Histochem Cytochem.

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“…Amplification by PCR was performed using standard Taq buffer (Promega) with the addition of 10% glycerol (30 cycles, denaturation 95°C, 1 min; annealing 50°C, 1 min; extension 72°C, 2 min). The primers for PCR were designed from the published sequence for rat atrial PAM (Stoffers, Green & Eipper, 1989) and corresponded to regions spanning the following functional domains of PAM: the peptidylglycine á_hydroxylating mono-oxygenase (PHM) domain (nucleotides 482-500 and 1012-1030, primers 1 and 2, respectively), the protease-sensitive domain (nucleotides 1490-1508 and 1770-1788, primers 4 and 5, respectively) and the transmembrane domain (nucleotides 2760-2778 and 3173-3192, primers 6 and 7, respectively). In addition, different species of PAM were distinguished using primer 7 and a primer (3) to nucleotides 1454-1473.…”

Section: Identification Of Pam Transcriptsmentioning

confidence: 99%

Regulation by gastric acid of the processing of progastrin‐derived peptides in rat antral mucosa

Macro

Bate

Varró

et al. 1997

The Journal of Physiology

184

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Inhibition of gastric acid secretion by proton pump inhibitors like omeprazole increases the synthesis and secretion of the pyloric antral hormone gastrin. We report here how omeprazole influences the conversion of the gastrin precursor to its final products, and the abundance of mRNAs encoding proteins associated with progastrin processing in rat antral mucosa. Progastrin processing was studied using a pulse–chase protocol in antral mucosa, incubated in vitro, from rats treated with omeprazole for up to 5 days. Labelled peptides were detected by on–line scintillation counting after immunoprecipitation and HPLC. The mRNAs encoding prohormone‐processing enzymes were identified by Northern blot, polymerase chain reaction or ribonuclease protection assay, and their cellular origins identified by immunocytochemistry. Cleavage of [3H]‐ and [35S]‐labelled progastrins at Arg‐94–95 or Arg‐57–58, and amidation at Phe‐92 were not influenced by pretreatment with omeprazole. In contrast, cleavage of G34 (the thirty‐four amino acid amidated gastrin) at Lys‐74–75 to give G17 (the seventeen amino acid amidated gastrin), and of G34–Gly to Gl7–Gly (G34 and G17 with COOH‐terminal glycine), was increased 3‐fold after treatment with omeprazole for either 1 or 5 days. Approximately 20% of newly synthesized amidated and Gly‐extended gastrins were secreted within 240 min of the labelling period in omeprazole‐treated samples, but secretion of labelled gastrins from control tissue was undetectable over a comparable period. The amidating enzyme, peptidylglycine α‐amidating mono‐oxygenase (PAM), the prohormone convertases PC1/3, PC2, PC5 and the PC2 chaperone 7B2 were localized to rat antral gastrin cells by immunocytochemistry. The relative abundance of mRNA species encoding 7B2, PC5 and PAM were unchanged after treatment with omeprazole for 5 days, whereas gastrin, PC1/3 and PC2 mRNAs are known to increase at this time. The main consequence of increased cleavage at Lys‐74–75 is the production of G17 and G17–Gly at the expense of G34 and G34–Gly, respectively. The latter have longer plasma half‐lives, and so their increased cleavage may serve to limit the rise in plasma gastrin concentration after inhibition of acid secretion. Changes in the abundance of mRNAs encoding prohormone‐processing enzymes cannot account for the rapidity of the changes in cleavage of progastrin at Lys residues after omeprazole.

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“…This could be explained if PAM molecules present in A-and B-cells were somehow different, so that each form could only be recognized by one of the antisera but not by the other. Such molecular differences between mouse A-and B-cell's PAM molecules could be due either to different posttranslational processing (e.g., proteolysis, glycosylation, phosphorylation), giving rise to conformational changes of the protein, or to alternative splicing, as previously reported in pancreatic carcinoma cells (Tateishi et al 1994) and in other tissues (Stoffers et al 1989(Stoffers et al ,1991Zhang et al 1997).…”

Section: Diversity Of Pam Enzymes In Mouse Endocrine Pancreatic Cell mentioning

confidence: 94%

Immunocytochemical Finding of the Amidating Enzymes in Mouse Pancreatic A-, B-, and D-cells

Garmendia

Rodriguez

Burrell

et al. 2002

J Histochem Cytochem.

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S U M M A R Y ␣ -Amidation is catalyzed by two enzymatic activities, peptidyl-glycine ␣ -hydroxylating mono-oxygenase (PHM) and peptidyl-␣ -hydroxyglycine ␣ -amidating lyase (PAL), denoted collectively as peptidyl-glycine ␣ -amidating mono-oxygenase (PAM), which also may include transmembrane and cytoplasmic domains. PAM is present in mammalian pancreas, where it appears to be abundant in the perinatal period. Nevertheless, there is no agreement on the cell type(s) that produces PAM or even on its presence in adults. In the present study we found PAM (PHM and cytoplasmic domain) immunoreactivity (IR) in A-, B-, and D-cells of adult mouse pancreas. In contrast to previous reports, PAM IR was found in B-cells of human and rat. Most of the B/D-cells were PAM immunoreactive, although with variable intensity, whereas less than half of A-cells displayed IR. Immunocytochemistry and Western blotting suggested the existence of different PAM molecules. Differences in the cellular distribution of IR for PAM domains were also observed. Whereas PHM-IR was extended throughout the cytoplasm in the three cell types, presumably in the secretory granules, IR for the cytoplasmic domain in A/D-cells was restricted to a juxtanuclear region, perhaps indicating its cleavage in Golgi areas. Although glucagon, insulin, and somatostatin are nonamidated, amidated peptides (glucagon-like peptide 1, adrenomedullin, proadrenomedullin N-terminal 20 peptide) were found in the three cell types.

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Alternative mRNA splicing generates multiple forms of peptidyl-glycine alpha-amidating monooxygenase in rat atrium.

Cited by 108 publications

References 29 publications

Distribution of peptidyl-glycine alpha-amidating mono-oxygenase (PAM) enzymes in normal human lung and in lung epithelial tumors.

Distribution of peptidyl-glycine alpha-amidating mono-oxygenase (PAM) enzymes in normal human lung and in lung epithelial tumors.

Regulation by gastric acid of the processing of progastrin‐derived peptides in rat antral mucosa

Immunocytochemical Finding of the Amidating Enzymes in Mouse Pancreatic A-, B-, and D-cells

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