Role of ribosomal protein L12 in gonococcal invasion of Hec1B cells.
Previous studies led to the development of a model of contact-induced enhanced gonococcal invasion of human reproductive cells that utilizes the lutropin receptor (LHr) as both the induction signal for conversion to this enhanced-gonococcal-invasion phenotype (Inv(+) GC) and as the specific Inv(+) GC uptake mechanism. This model proposes that gonococci express a surface feature that mimics human chorionic gonadotropin (hCG), the cognate ligand for LHr, and that this structure is responsible for the specific and productive interaction of GC with LHr. In this report, we identify a 13-kDa gonococcal protein with immunological similarities to hCG. The antiserum reactivity is specific since interaction with the 13-kDa gonococcal protein can be blocked by the addition of highly purified hCG. This gonococcal "hCG-like" protein, purified from two-dimensional gels and by immunoprecipitation, was determined by N-terminal sequencing to be the ribosomal protein L12. We present evidence that gonococcal L12 is membrane associated and surface exposed in gonococci, as shown by immunoblot analysis of soluble and insoluble gonococcal protein and antibody adsorption studies with fixed GC. Using highly purified recombinant gonococcal L12, we show that preincubation of Inv(-) GC with micromolar amounts of rL12 leads to a subsequent five- to eightfold increase in invasion of the human endometrial cell line, Hec1B. In addition, nanomolar concentrations of exogenous L12 inhibits gonococcal invasion to approximately 70% of the level in controls. Thus, we propose a novel cellular location for the gonococcal ribosomal protein L12 and concomitant function in LHr-mediated gonococcal invasion of human reproductive cells.
Spence
J M
JM
Department of Microbiology and Immunology, School of Medicine and Dentistry, University of Rochester, Rochester, New York 14642, USA.
Clark
V L
VL
D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D002460:Cell Line; D006063:Chorionic Gonadotropin / Q000032:analysis / Q000302:isolation & purification / Q000502:physiology*; D004717:Endometrium / Q000382:microbiology*; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000472:pathogenicity*; D011974:Receptors, LH / Q000502:physiology; D012269:Ribosomal Proteins / Q000502:physiology*
D006063:Chorionic Gonadotropin; D011974:Receptors, LH; D012269:Ribosomal Proteins; C016700:ribosomal protein L7-L12