Regulation of phagosomal acidification - Differential targeting of Na+/H+ exchangers, Na+/K+-ATPases, and vacuolar-type H+-ATPases

被引:120
作者
Hackam, DJ
Rotstein, OD
Zhang, WJ
Demaurex, N
Woodside, M
Tsai, O
Grinstein, S
机构
[1] UNIV TORONTO, DIV CELL BIOL, HOSP SICK CHILDREN, TORONTO, ON M5G 1X8, CANADA
[2] UNIV TORONTO, TORONTO HOSP, DEPT SURG, TORONTO, ON M5G 1X8, CANADA
关键词
D O I
10.1074/jbc.272.47.29810
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
Vacuolar type (V) ATPases are thought to be the main determinant of phagosomal acidification, In phagosomes containing mycobacteria, which ostensibly impair the delivery of V-ATPases to the phagosomal membrane, the pH would be expected to be near neutral, This prediction was tested by microfluorescence ratio imaging using macrophages from mice susceptible to mycobacterial infection, Although less acidic than their counterparts containing dead bacteria, phagosomes containing live Mycobacteria bovis were nearly 1 pH unit more acidic than the cytosol, suggesting the existence of alternate H+ transport mechanisms, We there fore investigated whether Na+/H+ exchange (NHE) contributes to phagosomal acidification, Immunoblotting, reverse transcriptase-polymerase chain reaction, and pharmacological studies indicated that NHE1 is the predominant isoform of the exchanger in macrophages. Fractionation revealed that NHE1 is incorporated into the phagosomal membrane, and measurements of pH indicated that it is functional in this location, Nevertheless, acidification of the lumen of phagosomes containing either latex beads or live M. bovis was insensitive to (3-methylsulfonyl-4-piperidinobenzoyl)-guanidine methanesulfonate, a potent inhibitor of NHE1, This may have been due to the absence of an appropriate lumen to cytosol Na+ gradient, because the phagosomal membrane was found to be devoid of Na+/K+ pumps, Unexpectedly, the acidification of M. bovis phagosomes was fully reversed by specific inhibitors of the vacuolar H+-ATPase, suggesting that ATPases are present only transiently or in reduced quantities in the phagosomal membrane, Alternatively, acid equivalents accumulated in endosomes by V-ATPases may be delivered to the mycobacterial phagosome by carrier vesicles devoid of ATPases.
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页码:29810 / 29820
页数:11
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