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1.
The vasopressin precursor is composed of 3 domains in line, namely vasopressin, MSEL-neurophysin and a glycopeptide referred to as copeptin, which are separated during the processing. In guinea pig neurohypophysis, the precursor is partially processed so that a two-domain fragment, MSEL-neurophysin--copeptin, can be found along with free MSEL-neurophysin adn copeptin. Guinea pig copeptin has been sequenced. It is a glycopeptide composed of 38 amino acid residues rather than the 39 found in other mammalian copeptins. Compared with other copeptins, that from guinea pig shows a few substitutions and the deletion of one acidic residue, probably in position 32. This deletion might be responsible for incomplete cleavage by the trypsin-like processing enzyme. 相似文献
2.
The guinea pig two-domain precursor of MSEL-neurophysin and copeptin has been passed through a trypsin-Sepharose column in order to mimic the enzyme processing by a membrane-bound endopeptidase. Only two cleavages were observed located in the inter-domain sequence (at Arg-94 and Arg-98), in contrast to several additional cleavages found when free neurophysin or copeptin is subjected to soluble trypsin. Because the physiological maturation involves a single cleavage at Arg-94, both local accessibility in the precursor and narrow specificity of the enzyme are implied in the processing. 相似文献
3.
C Lazure H S Saayman R J Naudé W Oelofsen M Chrétien 《International journal of peptide and protein research》1989,33(1):46-58
Mammalian neurohypophyseal hormones, oxytocin and vasopressin, are known to be synthesized as part of two larger precursors containing, respectively, a VLDV-neurophysin and a MSEL-neurophysin together with its associated glycopeptide. Starting from ostrich neurohypophyses, a "big" neurophysin was isolated and chemically characterized. Following sequence determination of the CNBr-derived fragments and of peptides obtained from trypsin and V8-protease digestion of the oxidized protein, this "big" neurophysin was found to contain an MSEL-neurophysin moiety (94 residues) still covalently associated with the COOH-terminal glycopeptide (38 residues, copeptin). This study demonstrates that the ostrich MSEL-neurophysin sequence closely resembles all known MSEL-neurophysin sequences and that, furthermore, it does not contain the single amino acid insertion shown previously in the ostrich VLDV-neurophysin. It is also shown that the stretch of amino acids, linking the MSEL-neurophysin and the copeptin, is clearly different from its mammalian homologues and lacks the Arg residue normally recognized by the cleaving enzyme. This study also demonstrates that the ostrich copeptin is more closely related to the amphibian copeptin sequence than to its mammalian homologue, leading to the hypothesis that two families of copeptin molecules might exist. Thus, the ostrich MSEL-neurophysin-copeptin molecule is the first "big" neurophysin reported in birds and, together with the guinea pig and amphibian homologues, represents the third example of partial or no neurophysin-copeptin cleavage. 相似文献
4.
Guinea pig MSEL-neurophysin. Sequence comparison of eight mammalian MSEL-neurophysins 总被引:1,自引:0,他引:1
M T Chauvet J Chauvet R Acher 《International journal of peptide and protein research》1987,30(5):676-682
The amino acid sequence of guinea pig MSEL-neurophysin has been determined using tryptic peptides derived from the performic acid-oxidized protein and staphylococcal proteinase peptides obtained from the reduced-carboxamidomethylated neurophysin. Guinea pig MSEL-neurophysin consists of a 93-residue polypeptide chain that shows 12 substitutions and 2 deletions when compared to bovine MSEL-neurophysin. It displays the highest number of variations among known mammalian MSEL-neurophysins. These variations are mainly found in the C-terminal region (residues 88-93). Moreover guinea pig MSEL-neurophysin, like rat homologous protein, exhibits substitutions in positions 2, 5, 29 and 81 and lacks an arginine in the penultimate position. Comparison between eight mammalian MSEL-neurophysins reveals a highly conserved region (residues 1 to 88) and a hypervariable region (residues 89 to 93/95). On the other hand the eight species examined are endowed with arginine vasopressin except pig, which has a lysine vasopressin. In the vasopressin-MSEL-neurophysin precursor, the hormonal moiety and the MSEL region of neurophysin (residues 1-9) are encoded by a common exon in ox, rat and man; it can be concluded that this exon is evolutionarily conservative in contrast to the one encoding the C-terminal region of MSEL-neurophysin. 相似文献
5.
A ‘big’ frog (Rana esculenta) neurophysin, encompassing sequences homologous to mammalian MSEL-neurophysin and copeptin, has been passed through a trypsin-Sepharose column in order to compare its conformation with that of the two-domain intermediate precursor isolated from guinea pig. Whereas the polypeptide possesses 8 arginine residues, only two cleavages were observed located in a putative inter-domain sequence (at Arg-94 and Arg-114). Because free vasotocin has been isolated from the frog, it is assumed that pro-vasotocin has a three-domain conformation similar to that of pro-vasopressin but processing in amphibians involves only one step rather than two steps as in mammals. 相似文献
6.
Structure, processing and evolution of the neurohypophysial hormone-neurophysin precursors 总被引:4,自引:0,他引:4
Neurohypophysial hormones and neurophysins are derived from common precursors processed during the axonal transport from the hypothalamus to the neurohypophysis. Two neurohormones, an oxytocin-like and a vasopressin-like, on one hand, two neurophysins, termed VLDV-and MSEL-neurophysins according to residues in positions 2, 3, 6 and 7, on the other, are usually found in vertebrate species. In contrast to placental mammals that have oxytocin and arginine vasopressin, marsupials have undergone a peculiar evolution. Two pressor peptides, lysipressin and vasopressin for American species, lysipressin and phenylpressin for Australian macropods, have been identified in individual glands and it is assumed that the primordial vasopressin gene has been duplicated in these lineages. On the other hand, the reptilian mesotocin is still present in Australian species instead of the mammalian oxytocin, while the North American opossum has both hormones and South American opossums have only oxytocin. The neurophysin domain of each precursor is encoded by 3 exons and different evolutionary rates have been found for the 3 corresponding parts of the protein. The central parts, encoded by the central exons, are evolutionarily very stable and nearly identical in the 2 neurophysins of a given species. Recurrent gene conversions have apparently linked the evolutions of the 2 precursor lineages. In mammals, the 3-domain precursor of vasopressin is processed in 2 stages: a first cleavage splitting off vasopressin and a second cleavage separating MSEL-neurophysin from copeptin. Two distinct enzymatic systems seem to be involved in these cleavages. Processing is usually complete at the level of the neurohypophysis, but an intermediate precursor encompassing MSEL -neurophysin and copeptin linked by an arginine residue has been characterized in guinea pig. In vitro processing of this intermediate through trypsin--Sepharose reveals cleavages only in the interdomain region. In non-mammalian tetrapods, such as birds and amphibians, mesotocin and vasotocin are associated with neurophysins in precursors similar to those found in mammals. However, processing of the vasotocin precursor seems to be different from the processing of the vasopressin precursor, with a single cleavage leading to the hormone release. 相似文献
7.
One-step processing of the amphibian vasotocin precursor: structure of a frog (Rana esculenta) "big" neurophysin 总被引:3,自引:0,他引:3
G Michel J Chauvet M T Chauvet R Acher 《Biochemical and biophysical research communications》1987,149(2):538-544
Vasotocin-associated neurophysin (MSEL-neurophysin) from the frog Rana esculenta has been isolated and sequenced through tryptic and staphylococcal proteinase peptides and cyanogen bromide fragments. This protein appears homologous to the mammalian vasopressin-associated neurophysin with a C-terminal glycopeptide extension homologous to the mammalian copeptin. In contrast to the two-step processing of mammalian vasopressin/MSEL-neurophysin/copeptin precursor, a single cleavage is therefore involved in the processing of the amphibian vasotocin/neurophysin precursor. It appears that the physiological release of the vasopressin-like hormone from the N-terminal end of the protein precursor is not dependent upon a previous trimming of the C-terminal copeptin-like moiety. 相似文献
8.
B Levy M T Chauvet J Chauvet R Acher 《International journal of peptide and protein research》1986,27(3):320-324
Vasopressin, MSEL-neurophysin and a glycopeptide, here referred to as copeptin, are three fragments of a common protein precursor processed during axonal transport from hypothalamus to neurohypophysis. Neurohormones and neurophysins purified from 7-9-month-old bovine foetuses have previously been shown to be identical with those found in the adult. Copeptin has now been isolated from 7-9-month and 3-month-old bovine foetuses and chemically characterized. It can be concluded from the nature of the three precursors that the same vasopressin gene is expressed in the adult and the 7-9-month-old foetus. 相似文献
9.
B Levy Y Rouille M T Chauvet J Chauvet R Acher 《International journal of peptide and protein research》1987,29(5):604-612
Neurohypophysial hormone precursors are small proteins processed into several fragments during axonal transport from hypothalamus to neurohypophysis. From 3-month-old fetal bovine pituitaries the three fragments of vasopressin precursor, arginine vasopressin, MSEL-neurophysin and copeptin, and the two fragments of oxytocin precursor, oxytocin and VLDV-neurophysin, have been isolated and characterized. These polypeptides are identical to those previously identified in the late fetus (7-9 months old) and in the adult. It is concluded that the same genes are expressed during fetal and adult lives, the vasopressin gene appearing roughly four times more active than the oxytocin gene in the early fetus. Vasotocin, mesotocin and additional neurophysin have not been detected in the early fetus. 相似文献
10.
The vasopressin precursor is composed of 3 domains, namely vasopressin, MSEL-neurophysin and a glycopeptide. Processing occurs during axonal transport from hypothalamus to neurohypophysis from which the 3 fragments can be isolated. The glycopeptide fragment of the rat vasopressin precursor has been purified and sequenced. Despite the fact that rat MSEL-neurophysin is shortened (93 residues instead of 95 for other mammals), rat glycopeptide has 39 residues, as do the other mammalian glycopeptides, suggesting a similar processing. Fifteen substitutions are however observed when compared to ox glycopeptide. The C-terminal part of MSEL-neurophysin (residues 77-93) and the glycopeptide are encoded by the same exon and the homologies when compared with their bovine counterparts are 58% and 62% respectively. In contrast, the central part of rat MSEL-neurophysin (residues 10-76), which is encoded by a separate exon, displays 96% of homology; vasopressin and the N-terminal part of MSEL-neurophysin (residues 1-9), encoded by a third exon, are nearly invariant. 相似文献
11.
These studies were aimed at an initial characterization of the human vasopressin precursor and the evaluation of factors leading to misfolding by the pathological 87STOP mutation. This mutation deletes the precursor's glycosylated copeptin segment, which has been considered unnecessary for folding, and the last seven neurophysin residues. We investigated the role in folding of the last seven neurophysin residues by comparing the properties of the 87STOP precursor and its derivative neurophysin with those of the corresponding wild-type proteins from which copeptin had been deleted, leading to the following conclusions. First, despite modulating effects on several protein properties, the last seven neurophysin residues do not make a significant net thermodynamic contribution to precursor folding; stabilities of the mutant and wild-type precursors to both guanidine denaturation and redox buffer unfolding are similar, as are in vitro folding rates. Second, the monomeric forms of both precursors are unstable and predicted to fold inefficiently at physiological pH and temperature, as evidenced by precursor behavior in redox buffers and by thermodynamic calculations. Third, both precursors are significantly less stable than the bovine oxytocin precursor. These results, together with earlier studies elsewhere of vasopressin precursor behavior within rat neurons, are shown to represent a self-consistent argument for a role for glycosylated copeptin in vasopressin precursor folding in vivo, copeptin most probably assisting refolding by facilitating interaction of misfolded monomers with the calnexin/calreticulin system. This hypothesis provides an explanation for the absence of copeptin in the more stable oxytocin precursor and suggests that the loss of copeptin contributes to 87STOP pathogenicity. Reported cell culture studies of rat precursor folding are also discussed in this context. Most generally, the results emphasize the significance of monomer stability in the folding pathways of oligomeric proteins. 相似文献
12.
M T Chauvet G Coffe J Chauvet R Acher 《Comptes rendus des séances de la Société de biologie et de ses filiales》1976,170(2):257-269
Neurohypophysial hormone-Neurophysin complexes have been prepared from posterior pituitary glands of Artiodactyla (ox, sheep, pig), Perissodactyla (horse) and Cetacea (whale), by fractionated salt precipitation. The components have been separated by molecular sieving in 0.2 M acetic acid and neurophysins have been purified by ion-exchange chromatography on DEAE-Sephadex A-50. Two types of neurophysins, MSEL-neurophysins and VLDV-neurophysins, can be distinguished according to the amino acid residues in positions 2, 3, 6 and 7. MSEL-neurophysins of sheep, ox and pig have been characterized by the amino acid sequence. Ovine and bovine MSEL-neurophysins are nearly identical (one substitution out of 95 residues) and porcine MSEL-neurophysin is very similar (four substitutions and an apparent 3-residue C-terminal deletion). The biological function of neurophysins might be the carriage of neurohypophysial hormones but in this respect, each type of neurophysin is not clearly specific for a given hormone. On the other hand, each neurophysin might share a common precursor with a neurohypophysial hormone, the two parts remaining associated after cleavage. However, in the sheep posterior pituitary gland, the molar proportions of the two types of neurophysins, oxytocin and arginine vasopressin, are not equal, MSEL-neurophysin being more abundant than the other components. If a common precursor exists, neurophysins and neurohypophysial hormones are not merely produced by a simple cleavage mechanism. 相似文献
13.
Tadashi Baba Henry B. Hoff Hiromi Nemoto Hang Lee Joanne Orth Yuji Arai George L. Gerton 《Molecular reproduction and development》1993,34(3):233-243
Spermatogenesis is a unique system of differentiation involving cellular remodeling and the biogenesis of sperm-specific organelles. To study the biogenesis of one such organelle, the acrosome, we have been examining the gene expression, biosynthesis, and targeting of specific acrosomal proteins during mammalian spermatogenesis. An acrosomal marker that we recently purified and began characterizing is acrogranin, a 67,000-molecular-weight glycoprotein originally isolated from guinea pig testes. This glycoprotein is detected in pachytene spermatocytes and is found later in the acrosomes of developing spermatids and sperm. Immunoblotting of several tissues and immunofluorescent localization in frozen sections of guinea pig testes suggested that acrogranin was a germ cell-specific glycoprotein that was expressed meiotically and post-meiotically. However, Northern blot analysis demonstrated that the mRNA for acrogranin was ubiquitously expressed in all guinea pig and mouse tissues examined. Furthermore, the primary structures of guinea pig and mouse acrogranins, deduced from the cDNA sequences, reveal that this glycoprotein is a cysteine-rich molecule with a motif that is tandemly repeated seven times, very similar to that of the human epithelin/granulin precursor. We conclude that guinea pig and mouse acrogranins are homologues of the precursor of the human and rat epithelin/granulin peptides previously demonstrated to have growth-modulating properties. © 1993 Wiley-Liss, Inc. 相似文献
14.
G Michel B Lévy M T Chauvet J Chauvet R Acher 《International journal of peptide and protein research》1990,36(3):302-307
Vasotocin-associated neurophysin (MSEL-neurophysin) has been purified from goose neurohypophysis through molecular sieving and high-pressure reverse-phase liquid chromatography (HPLC). The protein has a molecular mass (measured by SDS-polyacrylamide gel electrophoresis) of 17 kDa in contrast to 10 kDa found for the mammalian MSEL-neurophysins. Complete amino acid sequence (131 residues) has been determined mainly through tryptic or staphylococcal proteinase peptides derived from carboxyamidomethylated neurophysin, isolated by HPLC and microsequenced. N- and C-terminal sequences have been established by Edman degradation or action of carboxypeptidase Y, respectively, applied on the native protein. Goose MSEL-neurophysin is homologous to the two-domain "big" MSEL-neurophysin previously identified in the frog. It appears that in non-mammalian tetrapods, namely birds and amphibians, the proteolytic processing of the pro-vasotocin involves only one cleavage, releasing the hormone moiety and a "big" neurophysin with two domains homologous to mammalian MSEL-neurophysin and copeptin, respectively. Comparison of the avian protein with its mammalian and amphibian counterparts reveals that the first half of the polypeptide chain is evolutionarily much less variable than the second and that the goose protein resembles the frog protein much more than the mammalian one. 相似文献
15.
Species comparison of adenosine A1 receptors in isolated mammalian atrial and ventricular myocardium
Various species have been used as models to study the effects of adenosine (ADO) on atrial and ventricular myocardium, but few direct tissue comparisons between species have been made. This study further characterizes adenosine A(1) receptor binding, adenylate cyclase activity and direct and indirect A(1) receptor-mediated functional activity in atrial and ventricular tissue from Sprague-Dawley rats and Hartley guinea pigs. Rat right atria (RA) were found to be significantly more sensitive to cyclopentyladenosine (CPA), while guinea pig left atria (LA) were more sensitive to CPA. After the addition of isoproterenol (ISO), the reduction of CPA response in rat RA was significantly greater than in guinea pig; however, after ISO treatment, the guinea pig LA was more sensitive to CPA than the rat. Adenylate cyclase inhibition by CPA was significantly greater in atria and ventricles obtained from guinea pig than rat. In competition binding experiments, guinea pig RA had significantly more high affinity sites than rat, but the K(i)s were not significantly different. There were no significant differences between guinea pig LA and rat LA. Guinea pig ventricular tissue had fewer high affinity sites than rat without any differences in their K(i) values. In antagonist saturation experiments, the density and affinity of A(1) receptors in guinea pig cardiac membranes were significantly greater than in rat. Our results indicate definite species differences as well as tissue differences between rat and guinea pig. These differences must be considered when interpreting studies using rat and guinea pig tissue as models for cardiac function. 相似文献
16.
Competitive antagonists of bradykinin 总被引:35,自引:0,他引:35
The first sequence-related competitive inhibitors of the classic kinin in vitro (rat uterus guinea pig ileum) and in vivo (rat blood pressure) assays have been developed. Replacement of the proline residue at position 7 of bradykinin (BK) with a D-phenylalanine residue is the key modification which converts BK agonists into antagonists. [D-Phe7]-BK exhibits moderate (pA2 = 5.0) inhibition of BK activity on the guinea pig ileum but possesses weak BK-like myotropic activity on the isolated rat uterus and 2-4% of BK depressor potency in the rat blood pressure assay. The additional replacement of the phenylalanine residues at positions 5 and 8 of [D-Phe7]-BK with the isosteric beta-(2-thienyl)-alanine residue produces a potent antagonist of BK activity on the uterus (pA2 = 6.4), ileum (pA2 = 6.3), and in the rat blood pressure assay. The antagonism of BK action on smooth muscle is specific for kinins (BK, kallidin, Met-Lys-BK), but neither inhibitor antagonizes the smooth muscle activity of angiotensin or substance P. Inhibition is competitive and fully reversible. 相似文献
17.
Summary. The nucleotide sequence of cDNA that encodes hamster d-amino-acid oxidase (DAO) was determined. The cDNA consisted of 1,590 nucleotides and a poly(A) tail. It had an open reading
frame for a protein consisting of 346 amino acid residues. The number of the amino acid residues is the same as that of the
rat DAO. However, the hamster DAO has one residue more than mouse DAO and one residue less than human, pig, rabbit, and guinea
pig DAOs. Amino acid sequence of the hamster DAO was highly similar to those of mouse and rat DAOs: 89% and 88% of the amino
acid residues were identical between the hamster and mouse DAOs and between the hamster and rat DAOs, respectively. The homology
was slightly less between the hamster DAO and the human (81%), pig (78%), rabbit (78%), or guinea pig DAO (82%). It has been
proposed that the mouse and rat DAOs lack an amino acid residue corresponding to the 25th residue of the DAOs of other mammals.
However, a detailed comparison of the amino acid sequences as well as the underlying nucleotide sequences by inclusion of
the hamster ones revealed that the rodent DAOs does not lack the 25th, but the 27th residue.
Received January 16, 2002 Accepted June 20, 2002 Published online November 14, 2002
Authors' address: Dr. Ryuichi Konno, Department of Microbiology, Dokkyo University School of Medicine, Mibu, Tochigi 321-0293, Japan, Fax:
+81-282-86-5616 相似文献
18.
M.M.T. O''Hare S. Tenmoku L. Aakerlund L. Hilsted A. Johnsen T.W. Schwartz 《Regulatory peptides》1988,20(4):293-304
Neuropeptide Y (NPY) was isolated and characterised from acid-ethanol extracts of rabbit and guinea pig brain. In both instances the chromatographic purification was a two-step procedure of gel filtration followed by reverse-phase high-performance liquid chromatography. The amino acid sequence of rabbit and guinea pig NPY was found to be identical to human and rat NPY as deduced from the cDNA structures. With the exception of the porcine peptide, all mammalian NPYs characterised to date have a methionine residue in position 17. This methionine residue is readily oxidized as indicated by the high degree of spontaneous oxidation of peptides found in the rabbit and guinea pig brain extracts and in NPY extracted from a rat phaeochromocytoma cell line. It is concluded that NPY is among the most highly conserved peptides and that NPYs containing methionine in position 17 are prone to oxidation. 相似文献
19.
The lack of any information as to the origin of epidermal arachidonic acid, an important precursor of eicosanoids in the epidermis, prompted us to determine in vitro whether or not microsomal preparations from rat and guinea pig epidermis possess the delta 6 and delta 5 desaturase activities. The incubations were performed in parallel with microsomal preparations from liver of these animals where activities for these enzymes have previously been reported. The conversions of radioactive fatty acids were determined after methylation and separation of the 14C-fatty acid methyl esters by argentation thin layer chromatography. Data from these studies demonstrated that delta 5 desaturase activity is markedly lower in guinea pig liver than in rat liver. Interestingly, preparations from rat and guinea pig epidermis at all concentrations tested lacked the capacity to transform either linoleic acid into gammalinolenic acid or dihomogammalinolenic acid into arachidonic acid. This observation implies that arachidonic acid that is present in the epidermal phospholipids is biosynthesized elsewhere endogenously and transported to the epidermis for esterification into the phospholipids. The site of this biosynthesis is presumably the liver and the mode of transport to the epidermis remains to be determined. These studies indicate arachidonic acid per se as an essential fatty acid for the epidermis. 相似文献
20.
Suzuki O Mochida K Yamamoto Y Noguchi Y Takano K Matsuda J Ogura A 《Molecular reproduction and development》2002,62(3):335-342
The common alpha-subunit of glycoprotein hormones (CGalpha) is a core protein shared by follicle-stimulating hormone (FSH), luteinizing hormone (LH), and thyroid-stimulating hormone (TSH). In order to obtain a molecular basis for an efficient superovulation technique applicable to a wide range of animal species and to discuss the phylogenetic aspect based on molecules related to the reproductive system, we determined cDNA sequences of CGalpha in seven laboratory animals: the guinea pig, Mongolian gerbil, golden hamster, mastomys, Japanese field vole, the JF1 strain of Mus musculus molossinus, and rabbit. Comparison of the inferred CGalpha amino acid sequences of these animals and other mammals (human, mouse, rat, cow, pig, and sheep) showed that the signal peptides and the first ten residues at the N-terminus of the apoprotein were variable, while the rest of the apoproteins were highly conserved. In particular, all rodents had a leucine residue at the apoprotein N-terminus, except the guinea pig, which had a phenylalanine residue, as in the cow, pig, sheep, and rabbit. Phylogenetic trees constructed from amino acid sequences suggest a closer relationship between the guinea pig and artiodactyls than to rodents, confirming the taxonomic peculiarity of the guinea pig. 相似文献