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1.
The effects of glucagon-(1-21)-peptide on pancreatic exocrine secretion and plasma glucose levels were studied and compared with those of native glucagon in anesthetized dogs. Intravenous bolus administration of 1 nmol or 10 nmol/kg of glucagon-(1-21)-peptide evoked a significant inhibition of secretin-stimulated pancreatic juice secretion and protein output in a dose-dependent manner, as equimolar doses of glucagon did. Native glucagon induced an immediate and transient increase in pancreatic juice volume, which was followed by a significant inhibition. However, glucagon-(1-21)-peptide showed only the inhibitory action. Glucagon-(1-21)-peptide had no effect on plasma glucose levels even when a dose of 10 nmol/kg was given. The results suggest that the N-terminal amino-acid residues of glucagon play an important role in the inhibition of pancreatic exocrine secretion. 相似文献
2.
Serotonin- and ten peptide-immunoreactive (IR) cell types were identified in the digestive tract of sea bass (Dicentrarchus labrax L.) larvae of four morphofunctional phases ranging in age from hatching to 61 days. The sequence of appearance and location of endocrine cells during ontogenetic development of the larvae was determined. The differentiation of endocrine cells followed a distal-proximal gradient in the gut which paralleled the morphofunctional differentiation. Serotonin-IR cells were identified in the last portion of the digestive tract from phase I onwards and in the gastric region from phase III, before these regions were morphofunctionally differentiated; met-enkephalin-IR cells were identified from phase II onwards in both the differentiated rectum and the undifferentiated intestine; cholecystokinin (CCK)- and synthetic human gastrin-34-IR cells were located only in the intestine and first found in the undifferentiated intestine of phase II; human gastrin-17-, peptide YY (PYY)- and neuropeptide Y (NPY)-IR cells appeared in the intestine from phase II and in stomach in phase IV, when it showed gastric glands; pancreatic polypeptide (PP)- and glucagon-IR cells were observed in both intestine and stomach, but insulin- and somatostatin-IR cells only in stomach, from phase III, during which the intestine but not the stomach was differentiated. PP- and PYY-, PP- and glucagon-, and PYY- and glucagon-like immunoreactivities coexisted from their first appearance in some cells of the gut. 相似文献
3.
Previous studies have shown that pig intestine contains a 69 amino acid glucagon (glicentin) as well as a 37 amino acid glucagon (oxyntomodulin). In pig pancreas the 29 amino acid glucagon predominates. Since glucagon is thought to be expressed from a single gene in mammals, these differences in molecular forms indicate differential posttranslational processing of the glucagon precursor by different tissues. In the current study glucagon immunoreactivity (IR) was separately purified from dog pancreas, stomach mucosa and ileum mucosa. Purification and sequence analysis of the different tissue glucagons show that dog pancreas and stomach mucosa contain glucagon-29 while ileum mucosa contains glucagon-37 and glucagon-69. The latter is the major form present with glucagon-37 accounting for only 10-20% of the total ileum glucagon content. The N-terminal 32 amino acid portion of dog glucagon-69 differs at 6 sites from pig glucagon-69: RSLQDTEEKSRSFSAPQTEPLNDLDQMNEDKR... The C-terminal glucagon-37 is identical to pig oxyntomodulin. 相似文献
4.
Yunxia Wang Charles S. Bockman Sandor Lovas Peter W. Abel Richard F. Murphy J. Michael Conlon 《Journal of neurochemistry》1993,61(4):1231-1235
Abstract: γ-Preprotachykinin mRNA is the most abundant tachykinin mRNA in rat tissues, but the pathway of posttranslational processing of its translation product is unknown. An antiserum was raised against the synthetic peptide Asp-Ala-Gly-His-Gly-Gln-lle-Ser-His [neuropeptide γ-(1-9)-peptide, equivalent to γ-preprotachykinin-(72-80)-peptide], that showed <1% reactivity with intact neuropeptide γ and other tachykinins. Neuropeptide γ-(1-9)-peptide was detected by radioimmunoassay in relatively high concentrations in extracts of regions of rat brain and gastrointestinal tract. These concentrations correlated with (r = 0.99), but were significantly (p < 0.05) less than, the concentrations of neurokinin A-like immunoreactivity. The neuropeptide γ-(1-9)-like immunoreactivity in an extract of rat brain was eluted from a reverse-phase HPLC column in a single fraction with the same retention time as synthetic neuropeptide γ-(1 -9)-peptide. The synthetic peptide did not contract or relax isolated rat trachea, superior mesenteric artery, stomach fundus, or ileum, and the peptide did not affect the ability of neuropeptide 7 to contract the rat fundus. It is concluded that, in rat tissues, Lys70 -Arg71 in 7-preprotachykinin is a major site of posttranslational processing, but the resulting product, neuropeptide γ-(1-9)-peptide, is neither an agonist nor an antagonist at the neurokinin-2 (NK-2) receptor. 相似文献
5.
N Geary 《Peptides》1987,8(5):943-945
Rats were intraperitoneally injected with 115 or 230 nmol/kg pancreatic glucagon or equimolar doses of glucagon-(1-21) just before refeeding at the beginning of the dark phase after a 12 hr period of food deprivation. Glucagon-(1-21) and pancreatic glucagon have been reported to have similar visceral effects, but glucagon-(1-21) does not have pancreatic glucagon's metabolic effects. In this experiment, both doses of pancreatic glucagon, but neither dose of glucagon-(1-21), significantly decreased meal size. This indicates that the C-terminal octapeptide of pancreatic glucagon is necessary for its satiety effect, just as it is for its metabolic effects. 相似文献
6.
C Shaw D McKay C F Johnston D W Halton I Fairweather P Kitabgi K D Buchanan 《Peptides》1990,11(2):227-235
Posttranslational processing of the neurotensin/neuromedin N (NT/NN) precursor has been investigated in mouse brain and small intestine by means of region-specific radioimmunoassays coupled to chromatographic fractionations. In brain, total NT/NN immunoreactivity measured with a common C-terminal antiserum was 15.72 pmol/g. NT measured with an N-terminal antiserum was 9.74 pmol/g and NN measured with an N-terminal antiserum was 5.98 pmol/g. In small intestine, combined NT/NN immunoreactivity was 108.55 pmol/g, consisting of 66.37 pmol/g NT but only 0.96 pmol/g NN. Gel permeation chromatography and reverse phase HPLC revealed that the large discrepancy in the NT and NN values obtained in small intestinal extracts was due to the presence of a high molecular weight, hydrophobic peptide, which was reactive only with the common C-terminally directed antiserum. Pepsinization of this generated an immunoreactive peptide with similar chromatographic characteristics to NN. In mouse intestine, NN is only partially cleaved from the common NT/NN precursor, resulting in the presence of an N-terminally extended molecular species. This novel molecular species of neuromedin N may be the physiological mediator of certain peripheral biological effects hitherto attributed to neurotensin or neuromedin N. 相似文献
7.
Development of a non-extracted ''two-site'' immunoradiometric assay for corticotropin utilizing extreme amino- and carboxy-terminally directed antibodies.
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The development of a 'two-site' immunoradiometric assay (i.r.m.a.) for the direct estimation of human corticotropin-(1-39)-peptide in plasma is described. The assay is based on the simultaneous addition of 125I-labelled sheep anti-(N-terminal corticotropin) IgG (immunoglobulin G) antibodies and rabbit anti-(C-terminal corticotropin) antiserum to standards and unknowns (0.5 ml) followed by 18h incubation. The use of solid-phase reagents was avoided in order to minimize non-specific effects and the time required for reactants to reach equilibrium. Instead, the separation of corticotropin-bound from free labelled antibody is achieved by the addition of sheep anti-(rabbit IgG) antiserum, which precipitates bound labelled antibody by complex-formation with rabbit anti-corticotropin antibodies, which are also hormone-bound. Several 125I-labelled sheep anti-(N-terminal corticotropin) IgG preparations were assessed in the i.r.m.a. Although each was derived from antisera raised to a thyroglobulin conjugate of synthetic corticotropin-(1-24)-peptide (Synacthen), purification of immunoglobulins before iodination by selective immunoadsorption resulted in preparations with distinct specificities which demonstrated marked differences in binding to intact human corticotropin-(1-39)-peptide. These preparations are compared in combination with two rabbit anti-(C-terminal corticotropin) antisera. A 'two-site' assay based on the use of 125I-labelled sheep anti-[ corticotropin-(2-16)-peptide] IgG and rabbit anti-[corticotropin-(34-39)-peptide] antiserum was optimized, since steric inhibition of antibody binding was avoided with this combination and because the measurement of only intact human corticotropin-(1-39)-peptide and not fragments was assured by the use of terminal antibodies. This i.r.m.a. is characterized by rapid equilibration of reactants, a wide 'operating range' (the precision of dose estimates was less than 4% over the range 30-2200 pg/ml) and high sensitivity [8 pg of corticotropin/ml (95% confidence interval 3.7-12.0) (4 pg minimal detectable mass) can be detected directly in plasma]. 相似文献
8.
Neuroanatomical, electrophysiological and immunohistochemical techniques were used to describe correlations between soma morphology and electrophysiological properties in two groups of guinea-pig enteric neurones posing particular challenges. Lucifer Yellow-staining of 542 myenteric plexus neurones of duodenum revealed a great diversity of neuronal morphology. The distribution was: Dogiel Type I 27%, Dogiel Type II 54%, Stach Type IV 9%; 10% were unclassified. Correlations were sought in 59 of these cells between morphology and electrophysiological properties but no particular association was recognised. Dynorphin A(1-8)-like immunoreactivity (Dyn A(1-8)-IR) was found in up to 90% of identified submucous neurones of guinea-pig ileum. Of 62 S-neurones, 41 showed 'weak' and 19 had 'intense' Dyn A (1-8)-IR. There was no evidence of Dyn A(1-8)-IR in 2 S-neurones, nor in 8/8 AH-neurones. As for 11/16 vasoactive intestinal peptide- (VIP-) IR neurones, there was a strong correlation between the presence of 'weak' Dyn A(1-8)-IR and the occurrence of inhibitory (IPSPs) and slow excitatory synaptic potentials (EPSPs) (13/16 cells tested), which were never observed in neurones with 'intense' Dyn A(1-8)-IR (16/16) or neuropeptide Y (NPY)-IR (8/8). Similarly, 7/7 neurones with 'weak' Dyn A(1-8)-IR, but not those (7/7) with 'intense' Dyn A(1-8)-IR, hyperpolarised or showed a conductance change to noradrenaline. It was concluded that dynorphin A(1-8)-like-IR was contained in two populations of submucous neurone that are anatomically, immunohistochemically, electrophysiologically and pharmacologically distinct and closely related to those containing VIP and NPY. Furthermore, as in the myenteric plexus throughout the small intestine, opioid peptides are not expressed in Dogiel Type II cells. 相似文献
9.
Measurement and partial characterization of the multiple forms of neurokinin A-like immunoreactivity in carcinoid tumours 总被引:6,自引:0,他引:6
J M Conlon C F Deacon G Richter W E Schmidt F St?ckmann W Creutzfeldt 《Regulatory peptides》1986,13(2):183-196
An antiserum raised against neurokinin A has been used to demonstrate storage and release of neurokinin A-like immunoreactivity by carcinoid tumours. The antiserum showed reactivity towards members of the tachykinin family of polypeptides in the order: neurokinin A greater than eledoisin greater than neurokinin B greater than kassinin greater than substance P greater than physalaemin but the magnitude of the cross-reactivity with substance P and physalaemin was less than 1% of that of neurokinin A. A sensitive (IC50 238 fmol/ml; minimum detectable concentration, 9 fmol/ml) radioimmunoassay was set up using this antiserum. Extracts of metastatic tumour tissue from four patients with a primary carcinoid tumour in the midgut contained both neurokinin A-like immunoreactivity (NKA-LI) and substance P-like immunoreactivity (SP-LI). The concentrations (pmol/g wet weight) of NKA-LI and SP-LI in the tumours were: patient A 210, 201; patient B 2276, 6849; patient C 1198, 834 and patient D 424, 379. Analysis of the tumour extracts by reverse phase HPLC indicated that the NKA-LI was heterogeneous. Under two different conditions of chromatography, one component was eluted with the same retention time as neurokinin A. Two further components were more hydrophobic than neurokinin A but were not eluted with the retention time of neurokinin B. Analysis of these components by gel filtration indicated a molecular weight in the 3000-4000 range suggesting that they may be related to neuropeptide K, an N-terminally extended form of neurokinin A. NKA-LI and SP-LI were undetectable in the plasma of patients A and D but were elevated in patient B (NKA-LI 1005 +/- 114; SP-LI 345 +/- 85 fmol/ml) and patient C (NKA-LI 80 +/- 31; SP-LI 21 +/- 13 fmol/ml). 相似文献
10.
Louise Vedtofte Thóra B B?dvarsdóttir Allan E Karlsen R Scott Heller 《The journal of histochemistry and cytochemistry》2007,55(1):97-104
The desert gerbil Psammomys obesus, an established model of type 2 diabetes (T2D), has previously been shown to lack pancreatic and duodenal homeobox gene 1 (Pdx-1) expression. Pdx-1 deficiency leads to pancreas agenesis in both mice and humans. We have therefore further examined the pancreas of P. obesus during embryonic development. Using Pdx-1 antisera raised against evolutionary conserved epitopes, we failed to detect Pdx-1 immunoreactivity at any time points. However, at E14.5, Nkx6.1 immunoreactivity marks the nuclei of all epithelial cells of the ventral and dorsal pancreatic buds and the only endocrine cell types found at this time point are glucagon and PYY. At E18.5 the pancreas is well branched and both glucagon- and ghrelin-positive cells are scattered or found in clusters, whereas insulin-positive cells are not found. At E22.5, the acini of the exocrine pancreas are starting to mature, and amylase and carboxypeptidase A immunoreactivity is found scattered and not in all acini. Ghrelin-, glucagon-, PYY-, gastrin-, somatostatin (SS)-, pancreatic polypeptide (PP)-, and insulin-immunoreactive cells are found scattered or in small groups within or lining the developing ductal epithelium as marked by cytokeratin 19. Using degenerate PCR, the P. obesus Neurogenin-3 (Ngn-3) gene was cloned. Nucleotide and amino acid sequences show high homology with known Ngn-3 sequences. Using specific antiserum, we can observe that Ngn-3-immunoreactive cells are rare at E14.5 but readily detectable at E18.5 and E22.5. In conclusion, despite the lack of detection of Pdx-1, the P. obesus pancreas develops similarly to Muridae species, and the Ngn-3 sequence and expression pattern is highly conserved in P. obesus. 相似文献
11.
The nucleotide sequence of cDNA encoding the common biosynthetic precursor of substance P, neurokinin A and neuropeptide K (beta-preprotachykinin) predicts that, in the human, the precursor contains a C-terminal flanking peptide of 19 amino acid residues [beta-preprotachykinin(111-129)-peptide]. Using an antiserum raised against synthetic human beta-preprotachykinin(117-126)-peptide in radioimmunoassay, we have demonstrated that an extract of a human neuroendocrine tumor of the adrenal medulla contained approximately equimolar concentrations of C-terminal preprotachykinin immunoreactivity (C-PPT-IR), substance P and neurokinin A. The C-terminal preprotachykinin flanking peptide was purified to homogeneity and its primary structure was determined. The amino acid sequence of the peptide, Ala-Leu-Asn-Ser-Val-Ala-Tyr-Glu-Arg-Ser-Ala-Met-Gln-Asn-Tyr-Glu, indicates identity with beta-preprotachykinin(111-126)-peptide. The data suggest that the C-terminal flanking peptide, like the tachykinins, is packed into secretory storage vesicles but the Arg127-Arg128-Arg129 residues in human beta-preprotachykinin are removed from the peptide by the action of endogenous processing enzyme(s). 相似文献
12.
Acidic extraction of the thymus from two day old rats followed by purification on Sephadex G-50 gelfiltration revealed the presence of atrial natriuretic peptide-like material (IR-ANP) by radioimmunoassay. Verification of the obtained immunoreactivity has been achieved by the use of two different types of antisera, i.e. two antisera directed against ANP (99-126), the other antiserum recognizing the N-terminal fragment (11-37) of the precursor ANP (1-126) molecule. In addition, reverse phase high performance liquid chromatography (RP-HPLC) and high performance gel permeation chromatography (HP-GPC) monitored by the three antisera have been employed for analysis of the extracted IR-ANP. In both systems the IR-ANP corresponded to the 15 kDa-ANP molecule (1-126). Furthermore, by using immunohistochemical techniques a distinct localization of the IR-ANP material could be demonstrated. The outer cortical area of the thymus, containing mostly lymphoid cells, showed extensive immunostaining with the three different antisera. The data reported here indicate that the rat thymus is a source of ANP. 相似文献
13.
Glucagon-(19-29) exerts a biphasic action on the liver plasma membrane Ca2+ pump which is mediated by G proteins 总被引:1,自引:0,他引:1
S Lotersztajn C Pavoine V Brechler B Roche M Dufour D Le-Nguyen D Bataille F Pecker 《The Journal of biological chemistry》1990,265(17):9876-9880
We have recently shown that nanomolar concentrations of glucagon-(19-29), which can derive from native glucagon by proteolytic cleavage of the dibasic doublet Arg17-Arg18, inhibit the Ca2+ pump in liver plasma membrane vesicles independently of adenylyl cyclase activation (Mallat, A., Pavoine, C., Dufour, M., Lotersztajn, S., Bataille, D., and Pecker, F. (1987) Nature 325, 620-622). We report here that the regulation of the Ca2+ pump by glucagon-(19-29) is dependent on guanine nucleotides. In the presence of 10 microM guanosine 5'-3-O-(thio) triphosphate (GTP gamma S) or 75 microM GTP, glucagon-(19-29) caused a biphasic regulation of the Ca2+ pump. ATP-dependent Ca2+ transport was inhibited in the presence of 10 pM to 1 nM glucagon-(19-29), while higher concentrations of the peptide (1-100 nM) reversed the inhibition caused by lower ones. GTP gamma S alone, at high concentrations (100 microM), reproduced the inhibitory effect of glucagon-(19-29) and induced a 40% inhibition of the basal activity of the Ca2+ pump which was reversed by low concentrations of glucagon-(19-29) (10 pM to 1 nM). Treatment of rats with cholera toxin resulted in a 70% increase in the basal activity of the Ca2+ pump, a loss of sensitivity to GTP gamma S and to the biphasic regulation by glucagon-(19-29). Treatment with pertussis toxin did not affect the response of the Ca2+ pump to GTP gamma S and glucagon-(19-29). We conclude that glucagon-(19-29) can exert a biphasic effect on the Ca2+ pump which is mediated by G protein(s) sensitive to cholera toxin. 相似文献
14.
Somatostatin in epithelial cells of intestinal mucosa is present primarily as somatostatin 28 总被引:2,自引:0,他引:2
Region-specific antisera to [Tyr14]-SS28(1-14) were used to identify cells containing immunoreactivity to the SS28(1-14) fragment of somatostatin 28 (SS28) in gastric and intestinal mucosal epithelium and in pancreatic islets by immunoperoxidase staining. Radioimmunoassay with iodinated [Tyr14]-SS28(1-14) identified one antiserum (F4) to SS28(1-14) that cross-reacted equally with SS28(1-12), SS28(1-14) and SS28. Two other antisera (F3 and F8) to SS28(1-14) did not cross-react with SS28(1-12) and showed insignificant cross-reactivity to SS28. Immunostaining results showed that F4 stained the same cells that reacted with antiserum AS-10, which is specific for the cyclic tetradecapeptide somatostatin, SS28(15-28). Antisera F3, F4, and F8 all reacted with islet D cells and with somatostatin cells in the antral mucosa. However, only antiserum F4 detected immunoreactivity in mucosal epithelial cells; F3 and F8 did not bind to these cells. After sections of intestine were exposed to trypsin, however, epithelial cells containing immunoreactivity to SS28(1-14) were detected in intestinal mucosa with antisera F3 and F8. These results were obtained for duodenum, jejunum, ileum, and colon, but most of the epithelial cells with immunoreactivity to SS28(1-14) were in the duodenum. Both radioimmunoassay and immunostaining results suggest that F3 and F8 bind to a region of SS28(1-14) that is unavailable to antibodies in the intact SS28 molecule.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
15.
Munc-18 associates with syntaxin and serves as a negative regulator of exocytosis in the pancreatic beta -cell 总被引:2,自引:0,他引:2
Zhang W Efanov A Yang SN Fried G Kolare S Brown H Zaitsev S Berggren PO Meister B 《The Journal of biological chemistry》2000,275(52):41521-41527
The Munc-18 protein (mammalian homologue of the unc-18 gene; also called nSec1 or rbSec1) has been identified as an essential component of the synaptic vesicle fusion protein complex. The cellular and subcellular localization and functional role of Munc-18 protein in pancreatic beta-cells was investigated. Subcellular fractionation of insulin-secreting HIT-T15 cells revealed a 67-kDa protein in both cytosol and membrane fractions. Immunohistochemistry showed punctate Munc-18 immunoreactivity in the cytoplasm of rat pancreatic islet cells. Direct double-labeling immunofluorescence histochemistry combined with confocal laser microscopy revealed the presence of Munc-18 immunoreactivity in insulin-, glucagon-, pancreatic polypeptide-, and somatostatin-containing cells. Syntaxin 1 immunoreactivity was detected in extracts of HIT-T15 cells, which were immunoprecipitated using Munc-18 antiserum, suggesting an intimate association of Munc-18 with syntaxin 1. Administration of Munc-18 peptide or Munc-18 antiserum to streptolysin O-permeabilized HIT-T15 cells resulted in significantly increased insulin release, but did not have any significant effect on voltage-gated Ca(2+) channel activity. The findings taken together show that the Munc-18 protein is present in insulin-secreting beta-cells and implicate Munc-18 as a negative regulator of the insulin secretory machinery via a mechanism that does not involve syntaxin-associated Ca(2+) channels. 相似文献
16.
Chang SY Cha HR Uematsu S Akira S Igarashi O Kiyono H Kweon MN 《Journal of immunology (Baltimore, Md. : 1950)》2008,180(3):1609-1618
Although the mucosal and the systemic immune compartments are structurally and functionally independent, they engage in cross-talk under specific conditions. To investigate this cross-talk, we vaccinated mice with tetanus toxoid together with cholera toxin with s.c. priming followed by intrarectal (IR) boosting. Interestingly, higher numbers of Ag-specific IgA and IgG Ab-secreting cells (ASCs) were detected in the lamina propria of the large intestine of mice vaccinated s.c.-IR. Ag-specific ASCs from the colon migrated to SDF-1alpha/CXCL12 and mucosae-associated epithelial chemokine/CCL28, suggesting that CXCR4(+) and/or CCR10(+) IgA ASCs found in the large intestine after s.c.-IR are of systemic origin. In the colonic patches-null mice, IgA ASCs in the large intestine were completely depleted. Furthermore, the accumulation of IgA ASCs in the colonic patches by inhibition of their migration with FTY720 revealed that colonic patches are the IgA class-switching site after s.c.-IR. Most interestingly, s.c.-IR induced numbers of Ag-specific IgA ASCs in the large intestine of TLR2(-/-), TLR4(-/-), MyD88(-/-), and TRIF(-/-) mice that were comparable with those of wild-type mice. Taken together, our results suggest the possibility that cross-talk could occur between the large intestine and the systemic immune compartments via the colonic patches without the assistance of innate immunity. 相似文献
17.
E K Frandsen L Thim A J Moody J Markussen 《The Journal of biological chemistry》1985,260(12):7581-7584
Glucagon-(1-21) was prepared fully synthetically as well as by carboxypeptidase A digestion of natural porcine glucagon. Neither of the two preparations had glucagon agonistic effects with regard to receptor binding or adenylate cyclase activation in purified rat liver plasma membranes. Nor did these preparations contain lipolytic activity in isolated free fat cells. A preliminary batch of glucagon-(1-21) prepared by carboxypeptidase A digestion did, however, contain 1-2% glucagon bioactivity. This activity was separated from glucagon-(1-21) by high-performance liquid chromatography and quantitatively recovered in four minor hind peaks which eluted close to but not in a position identical to the elution position of native glucagon. 相似文献
18.
S V Wu P Chew F J Ho J H Walsh H Wong T D Lee M T Davis J E Shively J R Reeve 《Biochemical and biophysical research communications》1989,161(1):69-74
A peptide identical in structure to the carboxyl-terminal flanking nonapeptide of rat progastrin, predicted by cDNA sequence, was synthesized. The synthetic peptide was used for production of a rabbit antiserum. This antiserum was used to develop a radioimmunoassay specific for rat carboxyl terminal flanking peptide. This assay was used to monitor the purification of immunoreactivity from rat antral extracts. Gel permeation, anion exchange and reverse phase chromatography steps resulted in a single absorbance peak associated with the carboxyl terminal flanking peptide immunoreactivity. The purified peptide eluted in the same position as the synthetic peptide during all three types of chromatography. This material was shown to be identical in mass to Ser-Ala-Glu-Glu-Glu-Asp-Gln-Tyr-Asn, the predicted sequence of the carboxyl terminal nonapeptide of rat progastrin. 相似文献
19.
High molecular weight Alzheimer's disease amyloid peptide immunoreactivity in human serum and CSF is an immunoglobulin G 总被引:8,自引:0,他引:8
W M Pardridge H V Vinters B L Miller W W Tourtellotte J B Eisenberg J Yang 《Biochemical and biophysical research communications》1987,145(1):241-248
A radioimmunoassay (RIA) was developed to detect the 4200 Dalton amyloid (A4) peptide or it's precursor (A4P) in human serum or cerebrospinal fluid (CSF). A synthetic peptide containing the first 28 amino acids of the 43 amino acid A4 peptide was covalently coupled to bovine thyroglobulin and a polyclonal antiserum in rabbits was prepared. This antiserum was specific for vascular amyloid and neuritic plaques in Alzheimer's disease brain as detected by immunoperoxidase. The synthetic peptide, which has a tyrosine at residue 10, was iodinated with chloramine T and [125I]iodine and was purified to homogeneity by C4 reverse phase high performance liquid chromatography (HPLC). Extraction of human serum over a C18 Sep Pak cartridge indicated immunoreactive A4 peptide was not detectable in human serum. Conversely, high molecular weight A4 peptide immunoreactivity was detectable in human serum, at a concentration of 8.9 +/- 1.2 pmol-eq./ml, and in human CSF, at a concentration of 0.25 +/- 0.01 pmol-eq./ml, giving a CSF/serum ratio of 3.2%. The immunoreactivity in human serum was nearly completely removed by affinity deletion of serum immunoglobulin G (IgG), but not by affinity removal of IgA or IgM. Serum immunoreactivity was decreased 90% in hypogammaglobulinemia, and was increased 83% in human cord serum. There was no statistical difference in serum A4 immunoreactivity in Alzheimer's serum or CSF. Serum immunoreactivity in Down's syndrome was increased 50%. These studies indicate the high molecular weight A4P immunoreactivity in human serum or CSF is an IgG. Whether the A4 precursor in Alzheimer's disease is, in fact, an IgG, or whether there is an antibody in human serum and CSF that cross reacts with the A4 precursor cannot be determined until the serum immunoreactivity is purified and structurally characterized. 相似文献
20.
J. C. Garaud R. Eloy A. J. Moody C. Stock J. F. Grenier 《Cell and tissue research》1980,213(1):121-136
Summary The distribution and cellular location of substances reacting with anti-glucagon or anti-glicentin sera, i.e., glucagon-like and glicentin-like immunoreactivities, were studied in the human digestive tract using the immunofluorescence and immunoperoxidase methods. Both types of immunoreactivity were (1) absent in the antrum, (2) abundant in cells located at the periphery of pancreatic islets, (3) unevenly present in cells scattered in the epithelium of the small intestinal mucosa, the glicentin-immunoreactive cells being particularly abundant in the ileum. In the pancreas, and, when simultaneously present, in the intestine, both glucagon and glicentin immunoreactivities were located in the same cells.The precise ultrastructural location of each immunoreactivity was readily made using colloidal gold and ferritin tracers on ultrathin sections of glutaraldehyde-osmium fixed and epoxy resin-embedded tissues. In the pancreas, both glucagon and glicentin immunoreactivities were found in the granules of the A-type cells; the glucagon immunoreactivity was only present in the core of the granule, whereas the glicentin immunoreactivity was found either in the peripheral halo only, or throughout the entire granule. In the small intestine, both immunoreactivities were located inside the granules of the L-type cells.Quantitative specificity tests suggested that the glucagon- and the glicentin-like substances of the pancreas differ from those found in the intestine.Work supported by INSERM, A.T.P. number: 167539 相似文献