PCR cloning and baculovirus expression of human lactoperoxidase and myeloperoxidase

Kouichirou Shin, Hirotoshi Hayasawa, Bo Lönnerdal

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Lactoperoxidase (LPO) and myeloperoxidase (MPO) have been identified previously in human milk. These peroxidases have antimicrobial activity and presumably contribute to the protective functions of milk. In this study, we amplified genes encoding LPO and MPO from human mammary gland cDNA by the polymerase chain reaction (PCR). These genes were expressed in a baculovirus-insect cell system. Peroxidase activity was observed in the culture supernatant of Tricoplusia ni cells infected with the recombinant viruses and the levels increased upon addition of δ-aminolevulinic acid. Purified recombinant human LPO and MPO, both with a molecular mass of about 80 kDa, showed properties similar to bovine LPO and human MPO, respectively, in terms of absorption spectrum, sensitivity to dapsone, specificity for chloride ions, and reactivity with anti-bovine LPO or anti-MPO antibodies. Our data suggest that this expression system is useful for studying the catalytic mechanism and biological significance of these human peroxidases. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)831-836
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume271
Issue number3
DOIs
StatePublished - May 19 2000

Fingerprint

Lactoperoxidase
Baculoviridae
Cloning
Polymerase chain reaction
Peroxidase
Organism Cloning
Polymerase Chain Reaction
Peroxidases
Dapsone
Aminolevulinic Acid
Gene encoding
Molecular mass
Human Milk
Human Mammary Glands
Viruses
Genes
Insects
Chlorides
Absorption spectra
Anti-Idiotypic Antibodies

Keywords

  • Baculovirus-insect cell system
  • cDNA
  • Lactoperoxidase
  • Myeloperoxidase
  • PCR cloning

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

PCR cloning and baculovirus expression of human lactoperoxidase and myeloperoxidase. / Shin, Kouichirou; Hayasawa, Hirotoshi; Lönnerdal, Bo.

In: Biochemical and Biophysical Research Communications, Vol. 271, No. 3, 19.05.2000, p. 831-836.

Research output: Contribution to journalArticle

Shin, Kouichirou ; Hayasawa, Hirotoshi ; Lönnerdal, Bo. / PCR cloning and baculovirus expression of human lactoperoxidase and myeloperoxidase. In: Biochemical and Biophysical Research Communications. 2000 ; Vol. 271, No. 3. pp. 831-836.
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