Subject: intact organlles

From: [email-masked] (Nancy Keller)
Date: 15 Jun 1995 08:13:06 -0700

Hello everyone,
Q.  We are interested in isolating intact organlles (aspergillus) for
purposes of measuring pH of internal contents.  Has anyone experience w/
this or suggestions of references?  Thanking you,
Nancy P. Keller
Dept. Plant Pathology and Microbiology
Texas A&M University
College Station, TX 77843-2132
409-845-0963 P
409-845-8463 F
[email-masked]

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From: [email-masked] (David H. Griffin)
Date: 15 Jun 1995 19:55:09 GMT

In article <[email-masked]>
[email-masked] (Nancy Keller) writes:

> Hello everyone,
> Q.  We are interested in isolating intact organelles (aspergillus) for

I think that most people have given up on trying to measure pH of
isolated cell contents because of the uncertainties of what happens
during the isolation process. There are some ways of measuring
intracellular pH of intact cells. I don't know whether the resolution
is good enough to relate to specific organelles, vis:

Pena, A., J. Ramirez, G. Rosas and M. Calahorra.  1995.  Proton pumping
and the internal pH of yeast cells, measured with pyranine introduced
by electroporation.  J. Bacteriol. 177: 1017-1022.

Imai, T. and T. Ohno.  1995.  Measurement of yeast intracellular pH by
image processing and the change it undergoes during growth phase.  J.
Biotechnol. 38: 165-172.

Lohmeier-Vogel, E. M., B. Hahn-HŠgerdal and H. J. Vogel.  1995. 
Phosphorus-31 and carbon-13 nuclear magnetic resonance studies of
glucose and xylose metabolism in Candida tropicalis cell suspensions. 
Appl. Environ. Microbiol. 61: 1414-1419.

Slayman, C. L., V. V. Moussatos and W. W. Webb.  1994.  Endosomal
accumulation of pH indicator dyes delivered as acetoxymethyl esters. 
J. Exp. Biol. 196: 419-438.

Rowe, S. M., W. J. Simpson and J. R. M. Hammond.  1994.  Intracellular
pH of yeast during brewery fermentation.  Lett Appl Microbiol 18:
135-137.

Pilatus, U. and D. Techel.  1991.  P-31-NMR-studies on intracellular pH
and metabolite concentrations in relation to the circadian-rhythm,
temperature and nutrition in Neurospora crassa.  Biochim. Biophys. A.
1091: 349-355.

Yamashiro, C. T., P. M. Kane, D. F. Wolczyk, R. A. Preston and T. H.
Stevens.  1990.  Role of vacuolar acidification in protein sorting and
zymogen activation: a genetic analysis of the yeast vacuolar
proton-traslocating ATPase.  Mol. Cell. Biol. 10: 3737-3749.

Legerton, T. L., K. Kanamori, R. L. Weiss and J. D. Roberts.  1983. 
Measurements of cytoplasmic and vacuolar pH in Neurospora using
nitrogen-15 nuclear magnetic resonance spectroscopy.  Biochemistry 22:
899-903.

Dave.

David H. Griffin
Department of Environmental & Forest Biology
College of Environmental Science and Forestry
350 Illick Hall
One Forestry Drive
Syracuse NY 13210-2788
e-mail: [email-masked]