Alterations in the protein composition of maturing phagosomes

A. Pitt, L. S. Mayorga, P. D. Stahl, A. L. Schwartz

Research output: Contribution to journalArticle

164 Scopus citations

Abstract

We investigated the protein composition of J774-E clone macrophage phagosomes isolated at different stages of phagolysosome biogenesis. Phagosomes formed by internalizing antibody-coated Staphylococcus aureus for 3 min followed by chase for 0, 4, 9, or 15 min were isolated by density gradient centrifugation. Enrichment and purity of the phagosome preparations were quantitated by radiolabeled ligand recovery, enzyme markers, and electron microscopy. One-dimensional SDS-PAGE analyses of the isolated phagosomes revealed virtually identical protein compositions. However, Western blot analyses with antibodies directed against selected proteins of known itineraries along the endocytic pathway demonstrated distinct differences in phagosome protein compositions. Accumulating within the maturing phagosome were the 31-kD subunit of the vacuolar proton pump, cathepsin D, β-glucuronidase, the cation dependent mannose 6-phosphate receptor, and LAMP-1. Decreasing within the maturing phagosome were the FcII receptor, the mannose receptor, and alpha-adaptin. These results indicate that although the macrophage phagosome's total protein composition changes little during phagolysosome formation, the maturing phagosome both receives and eliminates, possibly by protein recycling, specific membrane and sequestered proteins.

Original languageEnglish
Pages (from-to)1978-1983
Number of pages6
JournalJournal of Clinical Investigation
Volume90
Issue number5
DOIs
StatePublished - Jan 1 1992

Keywords

  • endocytosis
  • phagocytosis
  • protein sorting
  • vesicle fusion
  • vesicle maturation

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