Home LiteratureArticle Details
PMID: 3199069 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Isolation of a complementary DNA clone encoding a precursor to human eosinophil major basic protein.

The Journal of experimental medicine ·Vol. 168 ·No. 6 ·1988-12-01 ·Pages 2295-308

McGrogan M, Simonsen C, Scott R, Griffith J, Ellis N, Kennedy J, Campanelli D, Nathan C, Gabay J

Abstract

A 14-kD protein was purified from human PMNs and its NH2-terminal sequence was determined. Comparison of a portion of the NH2-terminal sequence of this protein to the recently reported NH2-terminal sequence of eosinophil major basic protein (MBP) showed them to be identical. To aid further characterization of the structural and functional properties of this molecule, we isolated from an HL-60 cDNA library a single class of cDNA clones whose sequence matched exactly the NH2-terminal amino acid sequence of the 14-kD polypeptide. Northern analysis of HL-60 cells suggests that MBP is constitutively expressed in HL-60 cells and is highly transcribed from a single copy gene. The sequence of the full-length cDNA clones predicts that MBP is synthesized as a 23-kD precursor form (pro-MBP) which is subsequently cleaved to release the mature 14-kD MBP. The putative pro-MBP has a predicted pI of 6.0, but both the charged and the hydrophobic residues are asymmetrically distributed, creating a bipolar molecule. The NH2-terminal half has a predicted pI of 3.7 and is hydrophilic, while the COOH-terminal half (corresponding to mature MBP) has a predicted pI of 11.1 and is hydrophobic.

MeSH Terms
Base Sequence Blood Proteins/genetics Blotting, Southern DNA Eosinophil Granule Proteins Eosinophils/metabolism Humans Molecular Sequence Data Protein Precursors/genetics Restriction Mapping Ribonucleases Sequence Homology, Nucleic Acid
Chemicals
Blood Proteins Eosinophil Granule Proteins Protein Precursors DNA Ribonucleases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
McGrogan M
Invitron Corporation, Redwood City, California 94063.
Simonsen C
Scott R
Griffith J
Ellis N
Kennedy J
Campanelli D
Nathan C
Gabay J
References (44)
44 references, click to expand
  1. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose.
    Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408-12 PMID: 4504350
  2. Acidic precursor revealed in human eosinophil granule major basic protein cDNA.
    J Exp Med. 1988 Oct 1;168(4):1493-8 PMID: 3171483
  3. Cationic proteins of human granulocytes. II. Separation of the cationic proteins of the granules of leukemic myeloid cells.
    Blood. 1974 Aug;44(2):235-46 PMID: 4211856
  4. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene.
    Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961-5 PMID: 1105573
  5. Comparative properties of the Charcot-Leyden crystal protein and the major basic protein from human eosinophils.
    J Clin Invest. 1976 Mar;57(3):633-40 PMID: 942977
  6. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.
    J Mol Biol. 1977 Jun 15;113(1):237-51 PMID: 881736
  7. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  8. Purification and characterization of a potent bactericidal and membrane active protein from the granules of human polymorphonuclear leukocytes.
    J Biol Chem. 1978 Apr 25;253(8):2664-72 PMID: 344320
  9. How do eucaryotic ribosomes select initiation regions in messenger RNA?
    Cell. 1978 Dec;15(4):1109-23 PMID: 215319
  10. Damage to schistosomula of Schistosoma mansoni induced directly by eosinophil major basic protein.
    J Immunol. 1979 Jan;122(1):221-9 PMID: 570202
  11. Inhibition of intractable nucleases with ribonucleoside--vanadyl complexes: isolation of messenger ribonucleic acid from resting lymphocytes.
    Biochemistry. 1979 Nov 13;18(23):5143-9 PMID: 497174
  12. Cytotoxic properties of the eosinophil major basic protein.
    J Immunol. 1979 Dec;123(6):2925-7 PMID: 501097
  13. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.
    Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201-5 PMID: 6159641
  14. Elevated levels of the eosinophil granule major basic protein in the sputum of patients with bronchial asthma.
    Mayo Clin Proc. 1981 Jun;56(6):345-53 PMID: 7230899
  15. Destruction of bloodstream forms of Trypanosoma cruzi by eosinophil granule major basic protein.
    Am J Trop Med Hyg. 1981 Jul;30(4):775-9 PMID: 7020455
  16. Organization and expression of eucaryotic split genes coding for proteins.
    Annu Rev Biochem. 1981;50:349-83 PMID: 6791577
  17. Possible role of flanking nucleotides in recognition of the AUG initiator codon by eukaryotic ribosomes.
    Nucleic Acids Res. 1981 Oct 24;9(20):5233-52 PMID: 7301588
  18. An immunofluorescent method for specific staining of eosinophil granule major basic protein.
    J Immunol Methods. 1981;47(2):227-38 PMID: 7033389
  19. A simple method for displaying the hydropathic character of a protein.
    J Mol Biol. 1982 May 5;157(1):105-32 PMID: 7108955
  20. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments.
    Gene. 1982 Oct;19(3):269-76 PMID: 6295880
  21. Localization of eosinophil granule major basic protein in human basophils.
    J Exp Med. 1983 Sep 1;158(3):946-61 PMID: 6350526
  22. Distinctive cationic proteins of the human eosinophil granule: major basic protein, eosinophil cationic protein, and eosinophil-derived neurotoxin.
    J Immunol. 1983 Dec;131(6):2977-82 PMID: 6644025
  23. Peptide antibiotic subtilin is synthesized via precursor proteins.
    Biochem Biophys Res Commun. 1983 Oct 31;116(2):751-8 PMID: 6418167
  24. A simple and very efficient method for generating cDNA libraries.
    Gene. 1983 Nov;25(2-3):263-9 PMID: 6198242
  25. Biological activity of recombinant human interleukin-2 produced in Escherichia coli.
    Science. 1984 Mar 30;223(4643):1412-4 PMID: 6367046
  26. Cationic antimicrobial proteins isolated from human neutrophil granulocytes in the presence of diisopropyl fluorophosphate.
    Infect Immun. 1984 Jul;45(1):29-35 PMID: 6376359
  27. Localization of a molecule immunochemically similar to eosinophil major basic protein in human placenta.
    J Exp Med. 1984 Jul 1;160(1):29-41 PMID: 6376683
  28. Molecular cloning, cDNA sequencing, and chemical synthesis of cecropin B from Hyalophora cecropia.
    Proc Natl Acad Sci U S A. 1985 Apr;82(8):2240-3 PMID: 3857578
  29. Defensins. Natural peptide antibiotics of human neutrophils.
    J Clin Invest. 1985 Oct;76(4):1427-35 PMID: 2997278
  30. Biochemical properties of the eosinophil cationic protein and demonstration of its biosynthesis in vitro in marrow cells from patients with an eosinophilia.
    Blood. 1986 Feb;67(2):498-503 PMID: 3942834
  31. The eosinophil and the pathophysiology of asthma.
    J Allergy Clin Immunol. 1986 Apr;77(4):527-37 PMID: 3514730
  32. Biochemical and functional similarities between human eosinophil-derived neurotoxin and eosinophil cationic protein: homology with ribonuclease.
    Proc Natl Acad Sci U S A. 1986 May;83(10):3146-50 PMID: 3458170
  33. Localization of human eosinophil granule major basic protein, eosinophil cationic protein, and eosinophil-derived neurotoxin by immunoelectron microscopy.
    Lab Invest. 1986 Jun;54(6):656-62 PMID: 3520144
  34. Subcellular location and properties of bactericidal factors from human neutrophils.
    J Exp Med. 1986 Nov 1;164(5):1407-21 PMID: 3772295
  35. The eosinophilic leukocyte: structure and function.
    Adv Immunol. 1986;39:177-253 PMID: 3538819
  36. Magainins, a class of antimicrobial peptides from Xenopus skin: isolation, characterization of two active forms, and partial cDNA sequence of a precursor.
    Proc Natl Acad Sci U S A. 1987 Aug;84(15):5449-53 PMID: 3299384
  37. Posttranslational processing of a human myeloid lysosomal protein, myeloperoxidase.
    Blood. 1987 Oct;70(4):1143-50 PMID: 2820530
  38. A 25-kDa NH2-terminal fragment carries all the antibacterial activities of the human neutrophil 60-kDa bactericidal/permeability-increasing protein.
    J Biol Chem. 1987 Nov 5;262(31):14891-4 PMID: 3667613
  39. Eosinophil granule cationic proteins: major basic protein is distinct from the smaller subunit of eosinophil peroxidase.
    J Leukoc Biol. 1988 Jan;43(1):1-4 PMID: 3422083
  40. Oxygen-independent bactericidal systems. Mechanisms and disorders.
    Hematol Oncol Clin North Am. 1988 Mar;2(1):159-69 PMID: 3279018
  41. Prepeptide sequence of epidermin, a ribosomally synthesized antibiotic with four sulphide-rings.
    Nature. 1988 May 19;333(6170):276-8 PMID: 2835685
  42. Structure and bactericidal activity of an antibiotic dodecapeptide purified from bovine neutrophils.
    J Biol Chem. 1988 Jul 15;263(20):9573-5 PMID: 3290210
  43. Biochemical and amino acid sequence analysis of human eosinophil granule major basic protein.
    J Biol Chem. 1988 Sep 5;263(25):12559-63 PMID: 3410852
  44. Physiochemical and biological properties of the major basic protein from guinea pig eosinophil granules.
    J Exp Med. 1974 Aug 1;140(2):313-32 PMID: 4846413
Article Info
Journal
The Journal of experimental medicine
Abbr.
J Exp Med
ISSN
0022-1007
DOI
10.1084/jem.168.6.2295
Published
1988-12-01
Pages
2295-308
Language
English
Region
United States
NLM ID
2985109R
PMCID
PMC2189145
Subset
IM
Grants
NIAID NIH HHS · AI-23807 · United States
NCI NIH HHS · CA-43610 · United States
External Links
PubMed source DOI: 10.1084/jem.168.6.2295 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2