Monoclonal antibody detection of CD46 clustering beneath Neisseria gonorrhoeae microcolonies.
CD46 (membrane cofactor protein), a complement-regulatory protein that participates in innate and acquired immunity, also serves as a receptor for viral and bacterial pathogens. CD46 isoforms terminate in one of two cytoplasmic tails, Cyt1 or Cyt2, which differ in signaling and trafficking properties. Dissecting the functions of the two cytoplasmic tails in these cellular processes has been hampered by the absence of specific reagents. Here we report the construction of Cyt1- and Cyt2-specific monoclonal antibodies (MAbs). These MAbs recognize unique epitopes within the tails and can be used for immunofluorescence microscopy, immunoblotting, and immunoprecipitation. Studies of Neisseria gonorrhoeae-infected cells with the CD46 tail MAbs demonstrate the differential recruitment of Cyt1 and Cyt2 to the cortical plaque.
Weyand
Nathan J
NJ
Department of Molecular Microbiology & Immunology, L220, Oregon Health and Science University, Portland, OR 97201-3098, USA. weyandn@ohsu.edu
Lee
Shaun W
SW
Higashi
Dustin L
DL
Cawley
Daniel
D
Yoshihara
Paul
P
So
Magdalene
M
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000378:metabolism*; D001422:Bacterial Adhesion / Q000276:immunology; D045744:Cell Line, Tumor; D016764:Cell Polarity / Q000276:immunology; D003593:Cytoplasm / Q000276:immunology; D004847:Epithelial Cells / Q000276:immunology / Q000378:metabolism; D006801:Humans; D051916:Membrane Cofactor Protein / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000276:immunology* / Q000302:isolation & purification* / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D051916:Membrane Cofactor Protein; D020033:Protein Isoforms