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. 1991 Nov 15;88(22):10059–10063. doi: 10.1073/pnas.88.22.10059

Primary structure and functional expression of a glutaminyl cyclase.

T Pohl 1, M Zimmer 1, K Mugele 1, J Spiess 1
PMCID: PMC52867  PMID: 1946423

Abstract

A glutaminyl cyclase (QC) that is probably involved in the biosynthesis of pyroglutamyl peptides such as gonadotropin-releasing hormone and thyrotropin-releasing hormone has been purified to homogeneity from bovine anterior pituitary. On the basis of N-terminal sequence analysis, a 2088-base-pair cDNA clone was isolated from a bovine anterior pituitary library. From the nucleotide sequence of this clone, the primary structure of a 330-residue protein and a preceding 31-residue prepropeptide sequence was deduced. By transfection of COS-7 monkey cells with a QC cDNA/pCDM8 vector construct, QC activity was expressed. Hybridization with mRNAs of various bovine tissues revealed expression of QC mainly in brain tissue.

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Selected References

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  1. Braas K. M., Stoffers D. A., Eipper B. A., May V. Tissue specific expression of rat peptidylglycine alpha-amidating monooxygenase activity and mRNA. Mol Endocrinol. 1989 Sep;3(9):1387–1398. doi: 10.1210/mend-3-9-1387. [DOI] [PubMed] [Google Scholar]
  2. Busby W. H., Jr, Quackenbush G. E., Humm J., Youngblood W. W., Kizer J. S. An enzyme(s) that converts glutaminyl-peptides into pyroglutamyl-peptides. Presence in pituitary, brain, adrenal medulla, and lymphocytes. J Biol Chem. 1987 Jun 25;262(18):8532–8536. [PubMed] [Google Scholar]
  3. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eipper B. A., Mains R. E., Glembotski C. C. Identification in pituitary tissue of a peptide alpha-amidation activity that acts on glycine-extended peptides and requires molecular oxygen, copper, and ascorbic acid. Proc Natl Acad Sci U S A. 1983 Aug;80(16):5144–5148. doi: 10.1073/pnas.80.16.5144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fischer W. H., Spiess J. Identification of a mammalian glutaminyl cyclase converting glutaminyl into pyroglutamyl peptides. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3628–3632. doi: 10.1073/pnas.84.11.3628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fling S. P., Gregerson D. S. Peptide and protein molecular weight determination by electrophoresis using a high-molarity tris buffer system without urea. Anal Biochem. 1986 May 15;155(1):83–88. doi: 10.1016/0003-2697(86)90228-9. [DOI] [PubMed] [Google Scholar]
  8. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Koger J. B., Humm J., Kizer J. S. Assay of glutaminylpeptide cyclase. Methods Enzymol. 1989;168:358–365. doi: 10.1016/0076-6879(89)68027-5. [DOI] [PubMed] [Google Scholar]
  10. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  11. Lapps W., Eng J., Stern A. S., Gubler U. Expression of porcine cholecystokinin cDNA in a murine neuroendocrine cell line. Proteolytic processing, sulfation, and regulated secretion of cholecystokinin peptides. J Biol Chem. 1988 Sep 15;263(26):13456–13462. [PubMed] [Google Scholar]
  12. Lathe R. Synthetic oligonucleotide probes deduced from amino acid sequence data. Theoretical and practical considerations. J Mol Biol. 1985 May 5;183(1):1–12. doi: 10.1016/0022-2836(85)90276-1. [DOI] [PubMed] [Google Scholar]
  13. Lechan R. M., Wu P., Jackson I. M., Wolf H., Cooperman S., Mandel G., Goodman R. H. Thyrotropin-releasing hormone precursor: characterization in rat brain. Science. 1986 Jan 10;231(4734):159–161. doi: 10.1126/science.3079917. [DOI] [PubMed] [Google Scholar]
  14. MESSER M., OTTESEN M. ISOLATION AND PROPERTIES OF GLUTAMINE CYCLOTRANSFERASE OF DRIED PAPAYA LATEX. Biochim Biophys Acta. 1964 Nov 22;92:409–411. doi: 10.1016/0926-6569(64)90204-4. [DOI] [PubMed] [Google Scholar]
  15. MacCumber M. W., Snyder S. H., Ross C. A. Carboxypeptidase E (enkephalin convertase): mRNA distribution in rat brain by in situ hybridization. J Neurosci. 1990 Aug;10(8):2850–2860. doi: 10.1523/JNEUROSCI.10-08-02850.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  17. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  18. Pelham H. R. The retention signal for soluble proteins of the endoplasmic reticulum. Trends Biochem Sci. 1990 Dec;15(12):483–486. doi: 10.1016/0968-0004(90)90303-s. [DOI] [PubMed] [Google Scholar]
  19. Richter K., Kawashima E., Egger R., Kreil G. Biosynthesis of thyrotropin releasing hormone in the skin of Xenopus laevis: partial sequence of the precursor deduced from cloned cDNA. EMBO J. 1984 Mar;3(3):617–621. doi: 10.1002/j.1460-2075.1984.tb01857.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Seeburg P. H., Adelman J. P. Characterization of cDNA for precursor of human luteinizing hormone releasing hormone. Nature. 1984 Oct 18;311(5987):666–668. doi: 10.1038/311666a0. [DOI] [PubMed] [Google Scholar]
  22. Seed B. An LFA-3 cDNA encodes a phospholipid-linked membrane protein homologous to its receptor CD2. 1987 Oct 29-Nov 4Nature. 329(6142):840–842. doi: 10.1038/329840a0. [DOI] [PubMed] [Google Scholar]
  23. Seidah N. G., Gaspar L., Mion P., Marcinkiewicz M., Mbikay M., Chrétien M. cDNA sequence of two distinct pituitary proteins homologous to Kex2 and furin gene products: tissue-specific mRNAs encoding candidates for pro-hormone processing proteinases. DNA Cell Biol. 1990 Jul-Aug;9(6):415–424. doi: 10.1089/dna.1990.9.415. [DOI] [PubMed] [Google Scholar]
  24. Thomas G., Thorne B. A., Hruby D. E. Gene transfer techniques to study neuropeptide processing. Annu Rev Physiol. 1988;50:323–332. doi: 10.1146/annurev.ph.50.030188.001543. [DOI] [PubMed] [Google Scholar]
  25. Vale W., Spiess J., Rivier C., Rivier J. Characterization of a 41-residue ovine hypothalamic peptide that stimulates secretion of corticotropin and beta-endorphin. Science. 1981 Sep 18;213(4514):1394–1397. doi: 10.1126/science.6267699. [DOI] [PubMed] [Google Scholar]
  26. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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