title: Isolation of a complementary DNA clone encoding a precursor to human eosinophil major basic protein.
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.
authors.affiliation: Invitron Corporation, Redwood City, California 94063.
authors.forename: M
authors.initials: M
authors.lastname: McGrogan
authors.forename: C
authors.initials: C
authors.lastname: Simonsen
authors.forename: R
authors.initials: R
authors.lastname: Scott
authors.forename: J
authors.initials: J
authors.lastname: Griffith
authors.forename: N
authors.initials: N
authors.lastname: Ellis
authors.forename: J
authors.initials: J
authors.lastname: Kennedy
authors.forename: D
authors.initials: D
authors.lastname: Campanelli
authors.forename: C
authors.initials: C
authors.lastname: Nathan
authors.forename: J
authors.initials: J
authors.lastname: Gabay
mesh_terms: D001483:Base Sequence; D001798:Blood Proteins / Q000235:genetics*; D015139:Blotting, Southern; D004247:DNA; D047091:Eosinophil Granule Proteins; D004804:Eosinophils / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000235:genetics; D015183:Restriction Mapping; D012260:Ribonucleases; D012689:Sequence Homology, Nucleic Acid
chemical_list: D001798:Blood Proteins; D047091:Eosinophil Granule Proteins; D011498:Protein Precursors; D004247:DNA; D012260:Ribonucleases