DOCSLIB.ORG

Further Characterization of Brain Cholecystokinin- Converting Enzymes (Arginine-Isoleucine Hydrolase/Arginine-Aspartate Hydrolase/Dipeptidases) STEVEN W

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Further Characterization of Brain Cholecystokinin- Converting Enzymes (Arginine-Isoleucine Hydrolase/Arginine-Aspartate Hydrolase/Dipeptidases) STEVEN W Proc. Nati. Acad. Sci. USA Vol. 77, No. 6, pp. 3669-3671, June 1980 Medical Sciences Further characterization of brain cholecystokinin- converting enzymes (arginine-isoleucine hydrolase/arginine-aspartate hydrolase/dipeptidases) STEVEN W. RYDER, EUGENE STRAUS, AND ROSALYN S. YALOW Solomon A. Berson Research Laboratory, Veterans Administration Medical Center, Bronx, New York 10468; and Department of Clinical Sciences, Albert Einstein College of Medicine at the Montefiore Hospital and Medical Center, Bronx, New York 10467 Contributed by Rosalyn S. Yalow, March 4, 1980 ABSTRACT The brain cholecystokinin-converting enzymes standard diluent containing the substrate. After 6 hr at 370C, that cleave intact cholecystokinin to its COOH-terminal dode- loo-M1 aliquots of the incubation mixture were removed, placed capeptide and octapeptide also cleave the synthetic dipeptides Arg-Ile (or Arg-Val or Arg-Leu) and Arg-Asp, respectively. Thus, in a boiling water bath for 5 min to inactivate the enzyme, and they are not hormone-specific enzymes but are bond-specific. then subjected to starch gel electrophoresis in order to deter- Ultracentrifuge studies demonstrate that there is Arg-Ile hy- mine the hormonal forms of the reaction products, as described drolase activity associated with a protein greater in molecular (1, 2). To establish reference markers for interpreting the starch weight than gamma globulin and that both Arg-Ile and Arg-Asp gel electrophoretic patterns, sulfated synthetic CCK8 (Squibb, hydrolase activities are associated with one or more proteins Princeton, NJ) and synthetic sulfated CCK12 (prepared by M. between albumin and gamma globulin in molecular weight. A. Ondetti and received through the courtesy of V. Mutt) were subjected to similar starch gel electrophoresis. We have previously described (1, 2) two partially purified en- The dipeptides L-arginyl-L-aspartic acid, L-arginyl-L-iso- zymes that are readily solubilized from extracts of mammalian leucine, L-arginyl-L-leucine, and L-arginyl-L-valine (Research brain and that convert porcine cholecystokinin (pCCK33) to Plus, Denville, NJ) were dissolved (30 mg/ml, 0.1 M barbital its COOH-terminal fragments, the dodecapeptide (CCK12) buffer) and 50 ,l of dipeptide solution was incubated at 37"C and the octapeptide (CCK8). These enzymes were proven to with 50 Ml of each of the enzyme solutions for assay of activity. differ from trypsin in size, temperature sensitivity, and substrate The dipeptide substrate solutions were also incubated in a specificity (1). Here, we report further characteristics of sub- similar manner with trypsin (Sigma; 5 mg/ml, 0.25 M phos- strate specificity and other physicochemical properties of these phate buffer at pH 7.5). At 3 and 24 hr, portions of the incu- enzymes. bation mixtures were placed in a boiling water bath and then applied to paper for chromatographic or electrophoretic MATERIALS AND METHODS analysis. Preparation and Partial Purification of Brain Enzymes. Separation of Arg-Ile, Arg-Leu, and Arg-Val from their re- Both crude bovine cortical extracts (bCE) and pooled Sephadex spective amino acid constituents was effected by paper chro- G-75 void volume eluates (G75 vv) of bCE were prepared ac- matography. Samples of the incubation mixtures (25 ,ul) were cording to published methods (2). applied to 3 MM Whatman paper and then subjected to de- In addition, ultracentrifugation was used to fractionate bCE. scending chromatography in a closed chamber with butanol/ The extract (0.7 ml), prepared as above, was layered above a acetic acid/water, 50:12:50 (vol/vol). After the solvent front discontinuous sucrose gradient (1 ml each of 50%, 25%, 22.5%, migrated 25-30 cm from the origin, the paper was dried and and 20% sucrose) in a 5-ml polyethylene tube and centrifuged stained with ninhydrin in order to identify the amino acid and at 40,000 rpm for 16 hr in a Spinco model L ultracentrifuge dipeptide regions. RF values were compared to those of refer- fitted with a Beckman swinging bucket rotor (SW 50.1). After ence amino acids and dipeptides run simultaneously with the centrifuge came to rest without braking, 0.5-ml samples samples of the incubates. were removed successively from the bottom of each tube. Each The products of enzyme incubations with Arg-Asp as sub- fraction was dialyzed for 1 hr at 40C against the standard dil- strate were separated by paper electrophoresis. Samples of the uent (0.1 M barbital buffer pH 8.6, containing 2.5 mg of bovine incubation mixture (25 ,l) were applied to paper strips and serum albumin per ml) and then studied for enzymatic activity. subjected to closed chamber electrophoresis (chamber buffer, As molecular weight markers, 125I-labeled gamma globulin and 0.1 M barbital; 10 V/cm). After 3 hr, the paper strips were dried bromphenol blue-stained albumin were layered in duplicate and stained with ninhydrin to identify the amino acid and di- tubes and centrifuged along with bCE. peptide positions. Reference amino acids and dipeptides were Substrates and Enzymatic Assay. Studies were performed electrophoresed concomitantly with the incubation aliquots. with both pCCK33 and synthetic dipeptides as substrates. Radioimmunoassay. Starch gel eluates were assayed for Both pCCK33 (KABI Diagnostica, Studsvik, Sweden) at a CCK peptides by using previously published methods (3, 4). concentration of 10 mg/ml and 125I-labeled pCCK33 (pCCK33 125I-Labeled gastrin-17 (porcine heptadecapeptide gastrin was used for labeling was a gift of Victor Mutt, Karolinska Institute, a gift from R. A. Gregory through the courtesy of Morton I. Stockholm, and was received through the Gastrointestinal Grossman, Wadsworth Veterans Administration Medical Hormone Research Service of the National Institute of Arthritis, Center) was used as tracer and synthetic CCK-8, as standard. Metabolism and Digestive Diseases, Bethesda, MD) were used Rabbit B antiserum crossreacts identically on a molar basis with as substrates. For enzyme assay, 0.4 ml of bCE or pooled G75 CCK33, CCK12, and CCK8. The minimal detectable concen- vv or ultracentrifuged sucrose gradient fraction was added to tration of CCK8 in this system is 5 pg/mI. The publication costs of this article were defrayed in part by page Abbreviations: pCCK33, 33 amino acid porcine cholecystokinin; charge payment. This article must therefore be hereby marked "ad- CCK12, COOH-terminal dodecapeptide; CCK8, COOH-terminal vertisement" in accordance with 18 U. S. C. §1734 solely to indicate octapeptide; bCE, bovine cortical extract; G75 vv, pooled Sephadex this fact. G-75 void volume eluates of bovine cortical extracts. 3669 Downloaded by guest on September 28, 2021 3670 Medical Sciences: Ryder et al. Froc. Natl. Acad. Sci. USA 77 (1980) Sucrose gradient fraction A another with a sedimentation velocity between that of gamma Albumin Bromphenol blue globulin and that of bromphenol blue-stained albumin (fraction 416 B). Shown in Fig. 1 are the starch gel electrophoretic patterns of '25I-labeled peptides produced by treatment of '25I-labeled 2k pCCK with fractions A and B and those obtained after treat- ° Sucrose gradient fraction B ment of the same preparation with bCE and with G75 vv. x621u Products of treatment with fraction B as well as with bCE in- cluded both CCK12 and CCK8, but CCK8 was not produced x 61 4 - by treatment with fraction A or with G75 vv. E z - G75 vv Production of CCK12 and of CCK8 requires cleavage of at- L2- Arg-Ile and Arg-Asp bonds, respectively. We therefore 0 tempted to determine whether the converting enzymes could : Whole cortical extract hydrolyze these same bonds in the synthetic dipeptides. Ul- tracentrifuge fractions A and B as well as bCE and G75 vv all were active in cleaving the Arg-Ile bond although trypsin failed to do so (Fig. 2). The same converting enzymes from brain that Origin Anode cleaved Arg-Ile bonds also cleaved Arg-Leu and Arg-Val bonds FIG. 1. Starch gel electrophoretic patterns of 1251-labeled (Fig. 3). Thus, the specificity of these enzymes appears to be pCCK33 after treatment with (top to bottom) sucrose gradient frac- directed to the covalent bond between arginine and a neutral tion A (larger than gamma globulin), sucrose gradient fraction B amino acid whether in the dipeptide alone or in intact chole- (between human serumglobulin);vG75albumin and gamma v, and cystokinin. We have not yet investigated their ability to cleave crude unfractionated bCE from which the various enzymes were partially purified. Marker peptides run on similar gels confirmed that these bonds in other peptides. intact pCCK33 remained at the site of application or, on occasion, The descending chromatography system used for the sepa- migrated slightly cathodally; CCK12 migrated just anodally to al- ration of the arginyl-neutral amino acid dipeptides from the bumin and CCK8 migrated anodally to CCK12 and cathodally to single amino acids after cleavage of the covalent bonds could bromphenol blue. not resolve arginine and aspartic acid from Arg-Asp. However, RESULTS paper electrophoresis readily did so (Fig. 4). Treatment of In confirmation of our earlier findings (1, 2), we observed that pCCK33 with fraction A or G75 vv did not produce CCK8, and (i) bCE converted both unlabeled and '2I-labeled pCCK33 to Arg-Asp was not cleaved by either of these fractions. Although fragments with starch gel electrophoretic mobilities resembling Arg-Asp was not cleaved by trypsin, it was cleaved by treatment those of both CCK12 and CCK8 and (U) after treatment with with bCE and with the lower molecular weight proteins in ul- G75 vv, only CCK12 was produced. Enzymatic activity in the tracentrifuge fraction B. Thus, included in whole cortical ex- dialyzed sucrose gradient eluates peaked in the 50% and tracts are enzymes that have a molecular weight roughly be- 20%-22.5% sucrose fractions whereas radioactivity associated tween that of albumin and that gamma globulin and that can with 125I-labeled gamma globulin peaked in the 22.5%-25% cleave the Arg-Asp bond whether in the form of the free di- sucrose fractions.
Load more

Recommended publications
  • Is Chromogranin a Prohormone? Had Only a Fraction of the Pancreastatin Potency of the Peptide Characterized by Lee E
    ~N...:.AT.;:...U=--R.;:...E_V_O_L_.~32_5 _22_J_A_NU_A_R_Y_l_98_7 _________ NEWS ANDVIEWS-------------------'--301 Peptide function small percentage of plasma chromogranin was processed to a pancreastatin-like molecule, or if native chromogranin A Is chromogranin a prohormone? had only a fraction of the pancreastatin potency of the peptide characterized by Lee E. Eiden Tatemoto et at .. It is clearly necessary to establish that the molecule isolated by IN THE 4 December issue of Nature!, K. terminal processing signal for the forma- Tatemoto et at. is indeed the substance Tatemoto and co-workers report the tion of pancreastatin from a chromogranin found at beta-cell and D-cell receptors in sequence of a newly discovered putative A-like precursor in the rat. vivo. It will be of special interest to deter- hormone, pancreastatin, which as its There is chromogranin, as well as mine if amidation is necessary for bio­ name implies inhibits insulin (and soma­ pancreastatin, immunoreactivity in the logical activity, because amidation of tostatin) secretion from the endocrine pancreas of several species, and both poly- other biologically active peptides such as pancreas. There is a striking structural peptides are present in the brain"!', consis- enkephalins seems to oc-cur differentially similarity between pancreastatin and part tent with a precursor-product relationship in central and autonomic nervous tissue, of bovine chromogranin A, whose se­ between chromogranin A and pancreas- and this differential post-translational quence was recently reported in Nature by tatin. Whether or not pancreastatin is gen- modification may be relevant to different, myself and collaborators, and independ­ erated from chromogranin A or a chromo- as yet undiscovered hormonal functions of ently in the EMBO Journal by Benedum granin A-like precursor must await the these molecules.
    [Show full text]
  • Thiol-Disulfide Exchange in Human Growth Hormone Saradha Chandrasekhar Purdue University
    Purdue University Purdue e-Pubs Open Access Dissertations Theses and Dissertations January 2015 Thiol-Disulfide Exchange in Human Growth Hormone Saradha Chandrasekhar Purdue University Follow this and additional works at: https://docs.lib.purdue.edu/open_access_dissertations Recommended Citation Chandrasekhar, Saradha, "Thiol-Disulfide Exchange in Human Growth Hormone" (2015). Open Access Dissertations. 1449. https://docs.lib.purdue.edu/open_access_dissertations/1449 This document has been made available through Purdue e-Pubs, a service of the Purdue University Libraries. Please contact [email protected] for additional information. i THIOL-DISULFIDE EXCHANGE IN HUMAN GROWTH HORMONE A Dissertation Submitted to the Faculty of Purdue University by Saradha Chandrasekhar In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy August 2015 Purdue University West Lafayette, Indiana ii To my parents Chandrasekhar and Visalakshi & To my fiancé Niranjan iii ACKNOWLEDGEMENTS I would like to thank Dr. Elizabeth M. Topp for her tremendous support, guidance and valuable suggestions throughout. The successful completion of my PhD program would not have been possible without her constant encouragement and enthusiasm. Through the last five years, I’ve learned so much as a graduate student in her lab. My thesis committee members: Dr. Stephen R. Byrn, Dr. Gregory T. Knipp and Dr. Weiguo A. Tao, thank you for your time and for all your valuable comments during my oral preliminary exam. I would also like to thank Dr. Fred Regnier for his suggestions with the work on human growth hormone. I am grateful to all my lab members and friends for their assistance and support. I would like to especially thank Dr.
    [Show full text]
  • A Process for Producing Human Growth Hormone
    Patentamt JEuropaischesEuropean Patent Office (in Publication number: 0 217 814 Office europeen des brevets B1 EUROPEAN PATENT SPECIFICATION Date of publication of patent specification: 16.05.90 int.ci.5: C 12 P 21/06, C 12 N 15/00 Application number: 86901361.5 Date of filing: 06.02.86 International application number: PCT/DK86/00014 International publication number: WO 86/04609 14.08.86 Gazette 86/18 A PROCESS FOR PRODUCING HUMAN GROWTH HORMONE. (§) Priority: 07.02.85 DK 556/85 Proprietor: NOVO-NORDISK A/S Novo Alle DK-2880 Bagsvaerd (DK) Date of publication of application: 15.04.87 Bulletin 87/16 Inventor: ANDERSEN, Henrik, Dalboge Ordrupvej 1 12 B Publication of the grant of the patent: DK-2920 Charlottenlund (DK) 16.05.90 Bulletin 90/20 Inventor: PEDERSEN, John Kajerodvaenge 35 DK-3460 Birkero /d (DK) Designated Contracting States: Inventor: CHRISTENSEN, Thorkild AT BE CH DE FR GB IT LI LU NL SE Bellisvej 55 DK-3450Allerod(DK) Inventor: HANSEN, Jorli, Winnie References cited: Kloverhoj20 EP-A-0020290 DK-2600Glostrup(DK) EP-A-0089626 Inventor: JESSEN, Torben, Ehlern WO-A-84/02351 Kalundborgvej 216 US-A-4543329 DK-4300Holbaek(DK) Chemical Abstracts, vol. 77, 1972, Abstract no. 2289q, Calluhan et al, Fed. Proc. Fed. Amer, Coc. Representative: von Kreisler, Alek, Dipl.-Chem. Exp. Biol. 1972 31(3), 1105-13 (Eng). etal 00 Deichmannhaus am Hauptbahnhof Chemical Abstracts, vol. 76, 1972, Abstract no. D-5000 Koln 1 (DE) 83215s, Lindley et al. Biochem J. 1972. 126(3), 683-5(Eng). O Note: Within nine months from the publication of the mention of the grant of the European patent, any person may give notice to the European Patent Office of opposition to the European patent granted.
    [Show full text]
  • Evidence That the Major Degradation Product of Glucose-Dependent Insulinotropic Polypeptide, GIP(3–42), Is a GIP Receptor Antagonist in Vivo
    525 Evidence that the major degradation product of glucose-dependent insulinotropic polypeptide, GIP(3–42), is a GIP receptor antagonist in vivo V A Gault, J C Parker, P Harriott1, P R Flatt and F P M O’Harte School of Biomedical Sciences, University of Ulster, Cromore Road, Coleraine, N Ireland BT52 1SA, UK 1Centre for Peptide and Protein Engineering, School of Biology and Biochemistry, The Queen’s University of Belfast, Medical Biology Centre, Belfast, N Ireland BT9 7BL, UK (Requests for offprints should be addressed to V A Gault; Email: [email protected]) Abstract The incretin hormone glucose-dependent insulinotropic was significantly less potent at stimulating insulin secretion polypeptide (GIP) is rapidly degraded in the circulation (1·9- to 3·2-fold; P<0·001), compared with native GIP by dipeptidyl peptidase IV forming the N-terminally and significantly inhibited GIP-stimulated (107 M) truncated peptide GIP(3–42). The present study exam- insulin secretion with maximal inhibition (48·86·2%; ined the biological activity of this abundant circulating P<0·001) observed at 107 M. In (ob/ob) mice, admin- fragment peptide to establish its possible role in GIP istration of GIP(3–42) significantly inhibited GIP- action. Human GIP and GIP(3–42) were synthesised stimulated insulin release (2·1-fold decrease; P<0·001) and by Fmoc solid-phase peptide synthesis, purified by exaggerated the glycaemic excursion (1·4-fold; P<0·001) HPLC and characterised by electrospray ionisation-mass induced by a conjoint glucose load. These data indicate spectrometry. In GIP receptor-transfected Chinese ham- that the N-terminally truncated GIP(3–42) fragment acts ster lung fibroblasts, GIP(3–42) dose dependently inhib- as a GIP receptor antagonist, moderating the insulin ited GIP-stimulated (107 M) cAMP production (up to secreting and metabolic actions of GIP in vivo.
    [Show full text]
  • V1-Vasopressin Antagonists
    Europaisches Patentamt ® 0 1 82 626 � European Patent Office © Publication number: Office europeen des brevets A2 © EUROPEAN PATENT APPLICATION © Application number: 85308325.1 © Int. CI.4: C07K 7/06 , C07K 7/1 6 , A61 K 37/02 (g)rs Date of filing: 15.11.85 ® Priority: 21.11.84 US 673828 © Applicant: SMITHKLINE BECKM AN CORPORATION P.O. Box 7929 1 Franklin Plaza © Date of publication of application: Philadelphia Pennsylvania 191 01 (US) 28.05.86 Bulletin 86/22 © Inventor: Ali, Fadia Elfehail © Designated Contracting States: 5 Green Acre Drive, N. AT BE CH DE FR GB IT LI LU NL SE Cherry Hill New Jersey 08003(US) Inventor: Huffman, William Francis 40 Crest Avenue Malvern Pennsylvania 19355(US) © Representative: Waters, David Martin, Dr. et al Smith Kline & French Laboratories Ltd. Patent De- partment Mundells Welwyn Garden City Hertfordshire AL7 1EY(GB) © V1 -vasopressin antagonists. Compounds having the formula: in which: or a pharmaceutically acceptable salt thereof, which have V,-vasopressin antagonist activity and which are useful in the treatment of hypertension, shock and cardiac insuffi- P is Phe, Ile, Phe(4'-Alk), Tyr or Tyr(Alk); ciency. X is a D or L isomer of Val, Nva, Leu, Ile, Pba, Phe, Phe(4'-Alk), Trp, Nle, Cha, Abu, Met, Chg, Tyr or Tyr(Alk); Y is Val, Ile, Abu, Ala, Chg, Gln, Lys, Cha, Thr, Nle, Phe, Leu or Gly; Z is a D or L isomer of Arg, Harg, Leu, Lys or Orn; A is Gly(NH,), Gly, Gly(NH-Alk), OH, NH, or NHAlk; and R is, each, hydrogen or methyl; This invention relates to cyclic peptides which have studied, the β,β-diethyl and β,β-cyclopentamethylene pro- vasopressin (VSP) antagonist activity.
    [Show full text]
  • Page 1 of 30 Food & Function
    Food & Function Accepted Manuscript This is an Accepted Manuscript, which has been through the Royal Society of Chemistry peer review process and has been accepted for publication. Accepted Manuscripts are published online shortly after acceptance, before technical editing, formatting and proof reading. Using this free service, authors can make their results available to the community, in citable form, before we publish the edited article. We will replace this Accepted Manuscript with the edited and formatted Advance Article as soon as it is available. You can find more information about Accepted Manuscripts in the Information for Authors. Please note that technical editing may introduce minor changes to the text and/or graphics, which may alter content. The journal’s standard Terms & Conditions and the Ethical guidelines still apply. In no event shall the Royal Society of Chemistry be held responsible for any errors or omissions in this Accepted Manuscript or any consequences arising from the use of any information it contains. www.rsc.org/foodfunction Page 1 of 30 Food & Function Novelty of work In vitro transport of β-CM7 is limited by rapid hydrolysis into three peptide metabolites that transport at variable rates. 3.0 * 2.5 * * 2.0 * 1.5 1.0 * Manuscript Ratio to basal CCK-8 secretion CCK-8 basal to Ratio 0.5 0.0 M M M M M M M M M M M M M 500 250 125 500 250 125 500 250 125 1000 1000 1000 Fig. 8. Secretion of cholecystokinin[26-33] (CCK-8) by STC-1 cells in response to decreasing Accepted concentrations of YL and β-CM7 , compared to HBSS (basal) control .
    [Show full text]
  • Effect of Homochirality of Dipeptide to Polymers' Degradation
    polymers Article Effect of Homochirality of Dipeptide to Polymers’ Degradation Xinqiang Xu 1,2, Fuyan He 1,2, Wenke Yang 1,2 and Jinshui Yao 1,2,* 1 School of Materials Science & Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250353, China; [email protected] (X.X.); [email protected] (F.H.); [email protected] (W.Y.) 2 Shandong Provincial Key Laboratory of Processing & Testing Technology of Glass and Functional Ceramics, Jinan 250353, China * Correspondence: [email protected] Received: 31 August 2020; Accepted: 18 September 2020; Published: 22 September 2020 Abstract: As natural polymer materials, proteins are readily biodegradable, interestingly, the synthetic polyamides (PAs) that are based on the same amide bonds (also called peptide bonds in proteins) are barely degradable. Whether did the chirality and configuration of the amino acids play an important role. By using different configuration of amino acids, 4 types of polyamide-imides (PAIs) containing dipeptides of LL, DL, LD, and DD configurations, respectively, were synthesized. It was found that the PAIs based on natural LL configuration of dipeptide structure are much more readily biodegradable than those based on non-natural LD, DL, and DD configuration of dipeptides. It was confirmed that the natural L-configuration of amino acids play a critical role in degradability of proteins. And it also suggested that different type and amount of peptide fragments can be introduced in polymer to create series of polymer materials that can be biodegraded at controllable speed. Keywords: chiral amino acids; dipeptide; polyamide-imides; biodegradability 1. Introduction With the growing problem of white pollution, degradable polymer materials are attracting more and more attention because polymer materials that can be degraded at controllable speed could be used in every kind of fields [1–6].
    [Show full text]
  • Gastrin-Releasing Peptide (Grp)/Bombesin-Like Immunoreactivity in the Neurons and Paraneurons of the Gut and Lung
    Biomedical Research 4 (1) 93-104, 1983 GASTRIN-RELEASING PEPTIDE (GRP)/BOMBESIN-LIKE IMMUNOREACTIVITY IN THE NEURONS AND PARANEURONS OF THE GUT AND LUNG TOSHIHIKO IWANAGA Department of Anatomy, Niigata University School of Medicine, Niigata 951, Japan ABSTRACT Gastrin-releasing peptide (GRP)/bombesin-like immunoreactivities in the gut and lung were investigated with an immunocytochemical method using antisera showing different specificity for GRP-related peptides and bombesin. GRP-like, but not bombesin-like, immunoreactivity in the gut of birds and frogs was contained in the endocrine cells, which were restricted mainly to the oxyntic mucosa. On the other hand, GRP immunoreactivity in the rat and swine gut was contained in the nerve fibers distributed in the gastric mucosa and in the submucosal and myenteric plex- uses of the gut wall. In the lungs of human fetuses, the endocrine cells, solitary or gathered in groups, were immunoreactive to the GRP antisera. These endocrine cells and nerve fibers in the gut and lung showed the same immunoreactivity to two GRP antisera, but not to a bombesin-specific antiserum. This suggests that the bombesin- like immunoreactivities reported in the previous studies may be due to the existence of GRP or GRP-related peptides. Electron microscopic immunocytochemistry revealed that the GRP-like immunoreactivity was localized in the cytoplasmic gran- ules of the pulmonary endocrine cells and in the dense-cored vesicles of the gut nerve fibers. Bombesin is a tetradecapeptide isolated from the resides in the C-terminal portion of the molecule skin of Bombina bombina and Bombina variegara (17). Moreover, a heptacosapeptide having (1).
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 8,097.590 B2 Pinel Et Al
    USO08097590B2 (12) United States Patent (10) Patent No.: US 8,097.590 B2 Pinel et al. (45) Date of Patent: Jan. 17, 2012 (54) o-MSH-ANTAGONIST DIPEPTIDE FOREIGN PATENT DOCUMENTS CONUGATES EP O 389 95.0 A1 10, 1990 EP O 669938 B1 9, 1995 (75) Inventors: Anne-Marie Pinel, Toulouse (FR): E. : 29: R 1928 Pascal Verdie, Saint Mathieu de Treviers EP 1174 437 A1 1, 2002 (FR); Pascaline Dubs, Montpellier (FR): FR 1327363 5, 1962 Jean Martinez, Caux (FR); Gilles FR 2805744 A1 9, 2001 Subra, Juvignac (FR) FR 2810 323 A1 12/2001 s GB 1 OOO 897 8, 1965 (73) Assignees: Centre National de la Recherche W. W. 2.95. A. 3E Scientifique (CNRS), Paris (FR): WO WO 98.25584 A1 6, 1998 Institut Europeen de Biologie W. W. 9.3. A. 358: Cellulaire, Ramonville St. Agne (FR): WO WO O2/O85925 A2 10, 2002 Universite de Montpellier I, WO WO 03/064.458 A2 8, 2003 Montpellier (FR); Universite de WO WOO3,O95474 A2 11/2003 Montpellier II, Montpellier (FR) WO WO 2004/099237 A1 11, 2004 WO WO 2004/110341 A2 12/2004 (*) Notice: Subject to any disclaimer, the term of this WO WO 2005,116068 A1 12/2005 patent is extended or adjusted under 35 OTHER PUBLICATIONS U.S.C. 154(b) by 1143 days. Stverteczky et al., Acta Chimica Academiae Scientiarum (21) Appl. No.: 11/596,286 Hungaricae, 1975, vol. 87, No. 3, pp. 269-283.* Baumann et al., “Specificity of O-chyrinotrypsin dipeptide Sub (22) PCT Filed: May 10, 2005 strates.” FEBS Letters, Jun.
    [Show full text]
  • United States Patent Office Patented June 19, 1973
    3,740,385 United States Patent Office Patented June 19, 1973 1. 2 R-His-Ser-Asp-Gly 3,740,385 !-- N-TERMINAL DERVATIVES OF SECRETN Thr-Phe-Thr-Ser-Glu-Leu-Ser-Arg Miguel Angel Ondetti, 265 Hamlin Road, Leu-Arg-Asp-Ser-Ala-Arg-Leu-Glu(NH2)- North Brunswick, N.J. 08902 No Drawing. Continuation-in-part of abandoned applica Arg-Leu-Leu-Glu(NH2)-Gly-Leu-Val-NH2 tion Ser. No. 827,962, May 26, 1969. This application J. May 7, 1970, Ser. No. 35,598 R1-secretin Int. C. C07c 103/52, C07g 7/00 U.S. C. 260-112.5 6 Claims wherein R1 is a carboxylic acid, an amino acid, or a 10 dipeptide. The above equation takes into consideration that the ABSTRACT OF THE DISCLOSURE proper protection of the required moieties is followed. Disclosed herein are novel N-terminal derivatives of In addition when an acid is utilized, the desired acid the peptide secretin, intermediates thereof and a process may be reacted with secretin according to the equation: for their preparation. The secretin derivative of this in 5 vention are useful as antacids in hyperacidic conditions R2-COX--Secretin->R-CO-Secretin of the stomach and duodenum. wherein R2 is lower alkyl (e.g., methyl, ethyl, isopropyl, RELATED APPLICATION hexyl), aryl (e.g., phenyl or naphthyl), substituted phenyl 20 (e.g., o-chlorophenyl, p-ethylphenyl, m-trifluoromethyl This application is a continuation-in-part of my co phenyl, m-nitrophenyl), aralkyl (e.g., benzyl or phen pending application Ser. No. 827,962 filed May 26, 1969, ethyl), lower alkoxy (e.g., methoxy, ethoxy, butyloxy or now abandoned.
    [Show full text]
  • Development of Dipeptide Transport in the Intestinal Mucosa of Rabbits
    Pediatr. Res. 16: 99-103 (1982) Development of Dipeptide Transport in the Intestinal Mucosa of Rabbits STEFAN0 GUANDALINI'32' AND ARMIDO RUBINO Child Health Institute, 2nd School of Medicine, University of Naples, Italy Summary other hand, they can also be hydrolyzed at the brush border membrane, with-subsequent entjof the released amino acids. The uptake of 0.5 mM glycyl-L-proline (Gly-Pro), a substrate We had previously shown (24) that both ways do occur also in which has affinity for the dipeptide specific carrier system and is in the small intestine of the newborn rabbit, the transport of intact not hydrolyzed in the brush border, was studied vitro in jejunum dipeptide being quantitatively approximately as important as the and ileum of rabbits from the 25th day of gestational age into other pathway. We thus moved to investigate, in the developing adulthood. For comparison, uptake of 0.5 mM glycine was studied rabbit, the carrier system for dipeptides, with these major aims: too. Results show that influx of Gly-Pro is present from the (1) provide a biologic characterization of the system as for sub- earliest age studied and show a steep prenatal increase and peaks straie specificity, ~a dependence and kinetic constants; (2) define f at birth, with values, in the newborn jejunum, of 45.2 3.3 pmoles/ its developmental pattern as compared to that of the uptake of a ghr (mean + S.E.). Thereafter, the uptake slowly declines toward free neutral amino acid, as to provide an ontogenetic basis for the the adult level of 3.8 f 0.5 pmoles/ghr.
    [Show full text]
  • Fish Derived Bio-Active Peptides and Their Metabolic Effects Balık Kaynaklı Biyo-Aktif Peptidler Ve Metabolik Etkileri
    http://www.egejfas.org Su Ürünleri Dergisi (2015 Ege J Fish Aqua Sci 32(4): 217-223 (2015) DOI: 10.12714/egejfas.2015.32.4.08 REVIEW DERLEME Fish derived bio-active peptides and their metabolic effects Balık kaynaklı biyo-aktif peptidler ve metabolik etkileri Can Altınelataman1* • Anna Torkova2 • Mikhail Tsentalovich2 1Ege University, Faculty of Fisheries, 35100, Bornova, İzmir, Turkey 2Federal State Institution, Federal Research Centre, Fundamentals of Biotechnology of the Russian Academy of Sciences, Biochemistry Institute of Russian Academy of Sciences, Moscow, Russia *Corresponding Author: [email protected] How to cite this paper: Altınelataman, C., Torkova, A., Tsentalovich, M., 2015. Fish derived bio-active peptides and their metabolic effects. Ege J Fish Aqua Sci 32(4): 217-223. doi: 10.12714/egejfas.2015.32.4.08 Özet: Diğer tüm organizmalar gibi, balıklar da, araştırmalarda antimikrobiyal, antihipertensif, antioksidan ve antitümör aktivitelerine odaklanılan bio-aktif bileşiklerin zengin kaynağıdırlar. Bunlar, sadece yüksek besin değeri nedeniyle değil, özellikle son yirmi yılda geliştirilen analitik yöntemlerle elde edilen peptidleri açısından da kullanılabilirler. Akuatik canlılar dünya üzerinde en fazla tür sayısına sahiptir, dolayısıyla bunlardan bio-aktif peptidlerin eldesi diğerlerine göre oldukça olasıdır ve bu, önceki çalışmalarda açıkça ortaya konmuştur. Doğal peptidler ve uzun zincirli polipeptidlerden uygun enzimatik yöntemlerle elde edilen peptidler tıp alanında yeni ufuklar açabilirler. Anahtar kelimeler: Balık, peptidler, bio-aktif, metabolik etki, sağlık Abstract: Like many other organisms, fishes are also rich sources of bio-active compounds which were well studied by research focused on their antimicrobial, antihypertensive, antioxidant and antitumor activities. They can be used not only for good nutritional value, but also for peptides obtained with analytic processes that were developed in last decades.
    [Show full text]