ISOLATION OF PNEUMOCYSTIS-CARINII GP120 BY FIBRONECTIN AFFINITY - EVIDENCE FOR MANGANESE DEPENDENCE

被引:11
作者
WISNIOWSKI, P [1 ]
PASULA, R [1 ]
MARTIN, WJ [1 ]
机构
[1] INDIANA UNIV,WISHARD MEM HOSP,SCH MED,DEPT INTERNAL MED,DIV PULM & CRIT CARE,INDIANAPOLIS,IN 46202
关键词
D O I
10.1165/ajrcmb.11.3.8086164
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Pneumocystis carinii is a major opportunistic lung pathogen and a leading cause of death among patients with the human immunodeficiency virus. Adherence of P. carinii to type I alveolar epithelial cells is essential for growth and replication and has been shown to be mediated in part by fibronectin (Fn). To better understand the mechanisms underlying this attachment, P. carinii-Fn interaction was characterized with respect to divalent and monovalent ion concentration and pH using an I-125-Fn binding assay to Il carinii. The results suggest that P. carinii has a receptor for Fn that was partially dependent on Ca2+, enhanced by Mn2+, and diminished somewhat by Mg2+. Additional data demonstrated that P carinii-Fn interaction was sensitive to ionic strength. The pH profile revealed that P. carinii-Fn interaction increased with decreasing pH. The results from the binding assay provided the rationale for a simple isolation of the Fn receptor from P. carinii using a Fn-affinity column involving nondenaturing conditions. The isolated receptor appeared highly purified by SDS-PAGE analysis, with apparent molecular weights of 114 to 118 kD and 66 kD. Western blot analysis indicated that this receptor was gp120, a major surface glycoprotein of P. carinii. Furthermore, the isolated receptor inhibited Fn binding to P. carinii. Finally, a monoclonal antibody raised against the affinity-purified gp120 blocked Fn binding to P. carinii.
引用
收藏
页码:262 / 269
页数:8
相关论文
共 33 条
  • [1] NEW RAT MODEL OF PNEUMOCYSTIS-CARINII INFECTION
    BARTLETT, MS
    FISHMAN, JA
    QUEENER, SF
    DURKIN, MM
    JAY, MA
    SMITH, JW
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1988, 26 (06) : 1100 - 1102
  • [2] GROWTH AND METABOLISM OF PNEUMOCYSTIS-CARINII IN AXENIC CULTURE
    CUSHION, MT
    EBBETS, D
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1990, 28 (06) : 1385 - 1394
  • [3] DANIEL WW, 1987, F ANAL HLTH SCI
  • [4] DEBLAS AL, 1983, METHOD ENZYMOL, V92, P36
  • [5] RIBOSOMAL-RNA SEQUENCE SHOWS PNEUMOCYSTIS-CARINII TO BE A MEMBER OF THE FUNGI
    EDMAN, JC
    KOVACS, JA
    MASUR, H
    SANTI, DV
    ELWOOD, HJ
    SOGIN, ML
    [J]. NATURE, 1988, 334 (6182) : 519 - 522
  • [6] RECEPTOR FUNCTIONS FOR THE INTEGRIN VLA-3 - FIBRONECTIN, COLLAGEN, AND LAMININ BINDING ARE DIFFERENTIALLY INFLUENCED BY ARG-GLY-ASP PEPTIDE AND BY DIVALENT-CATIONS
    ELICES, MJ
    URRY, LA
    HEMLER, ME
    [J]. JOURNAL OF CELL BIOLOGY, 1991, 112 (01) : 169 - 181
  • [7] GAILIT J, 1988, J BIOL CHEM, V263, P12927
  • [8] HOST SPECIES-SPECIFIC ANTIGENIC VARIATION OF A MANNOSYLATED SURFACE GLYCOPROTEIN OF PNEUMOCYSTIS-CARINII
    GIGLIOTTI, F
    [J]. JOURNAL OF INFECTIOUS DISEASES, 1992, 165 (02) : 329 - 336
  • [9] DETECTION OF PNEUMOCYSTIS-CARINII BY FLUORESCENT-ANTIBODY STAIN USING A COMBINATION OF 3 MONOCLONAL-ANTIBODIES
    GILL, VJ
    EVANS, G
    STOCK, F
    PARRILLO, JE
    MASUR, H
    KOVACS, JA
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1987, 25 (10) : 1837 - 1840
  • [10] HAMM EK, 1971, EXP MOL PATHOL, V14, P362