EFFECT OF PHOSPHOLIPASE-C ON CHOLESTEROL SOLUBILIZATION IN MODEL BILE - A CONCANAVALIN-A BINDING NUCLEATION-PROMOTING FACTOR FROM HUMAN GALLBLADDER BILE

被引:49
作者
PATTINSON, NR [1 ]
WILLIS, KE [1 ]
机构
[1] CHRISTCHURCH HOSP,CHRISTCHURCH SCH MED,GASTROENTEROL RES UNIT,CHRISTCHURCH,NEW ZEALAND
关键词
D O I
10.1016/0016-5085(91)90086-Z
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Human bile contains a phospholipase C activity. To examine its pathophysiological importance, the effect of phospholipase C on the dynamics of lipid solubilization and nucleation (cholesterol crystal formation) were investigated in model bile. Phospholipase C from gallbladder bile from patients with gallstones was partially purified by competitively eluting from a concanavalin A (con A)-Sepharose (Sigma, St. Louis, MO) column and incubating with Pronase (Calbiochem, Behring Diagnostics, La Jolla, CA). Phospholipase C activity was resistant to Pronase digestion. When this fraction (concentrated to half the original volume) was mixed with model bile (1:1, vol/vol), a transfer of cholesterol and phospholipid from the micellar to the vesicular phase and an accelerate nucleation time were found concomitant with phospholipid hydrolysis. These effects were prevented by inhibiting the phospholipase C activity with ethylenediaminetetraacetic acid. To confirm that the results were caused by phospholipase C activity and not some other nucleation-promoting factor within the biliary con A preparation, model bile was incubated with bacterial phospholipase C. An identical cascade of events to that found with the partially purified biliary enzyme was observed. Further purification of the con A-bound proteins on DEAE-Sephadex (Pharmacia, Uppsala, Sweden) did not resolve any separate nucleation-promoting activity to that associated with phospholipase C activity. In conclusion, this study has identified phospholipase C as a/the con A nucleation-promoting activity in human gallbladder bile and has characterized a possible molecular mechanism by which cholesterol nucleation is stimulated by this fraction. © 1991.
引用
收藏
页码:1339 / 1344
页数:6
相关论文
共 26 条
  • [1] BURNSTEIN MJ, 1983, GASTROENTEROLOGY, V85, P801
  • [2] MODIFICATION BY DIACYLGLYCEROL OF THE STRUCTURE AND INTERACTION OF VARIOUS PHOSPHOLIPID-BILAYER MEMBRANES
    DAS, S
    RAND, RP
    [J]. BIOCHEMISTRY, 1986, 25 (10) : 2882 - 2889
  • [3] DUDLEY MA, 1986, GASTROENTEROLOGY, V90, P1722
  • [4] EFFECT OF MUCOUS GLYCOPROTEIN ON NUCLEATION TIME OF HUMAN BILE
    GALLINGER, S
    TAYLOR, RD
    HARVEY, PRC
    PETRUNKA, CN
    STRASBERG, SM
    [J]. GASTROENTEROLOGY, 1985, 89 (03) : 648 - 658
  • [5] GROEN AK, 1989, J LIPID RES, V30, P51
  • [6] CHOLESTEROL NUCLEATION-INFLUENCING ACTIVITY IN T-TUBE BILE
    GROEN, AK
    STOUT, JPJ
    DRAPERS, JAG
    HOEK, FJ
    GRIJM, R
    TYTGAT, GNJ
    [J]. HEPATOLOGY, 1988, 8 (02) : 347 - 352
  • [7] AN APPRAISAL OF THE ROLE OF BILIARY PHOSPHOLIPASES IN THE PATHOGENESIS OF GALLSTONE DISEASE
    GROEN, AK
    NOORDAM, C
    DRAPERS, JAG
    EGBERS, P
    HOEK, FJ
    TYTGAT, GNJ
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1989, 1006 (02) : 179 - 182
  • [8] RAPID VESICLE FORMATION AND AGGREGATION IN ABNORMAL HUMAN BILES - A TIME-LAPSE VIDEO-ENHANCED CONTRAST MICROSCOPY STUDY
    HALPERN, Z
    DUDLEY, MA
    KIBE, A
    LYNN, MP
    BREUER, AC
    HOLZBACH, RT
    [J]. GASTROENTEROLOGY, 1986, 90 (04) : 875 - 885
  • [9] HALPERN Z, 1986, J LIPID RES, V27, P295
  • [10] HOLAN KR, 1979, GASTROENTEROLOGY, V77, P611