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(Received for publication, April 4,
1994; and in revised form, September 13, 1994) From the
According to current concepts, new peroxisomes are formed by
division of pre-existing peroxisomes or by budding from a peroxisomal
reticulum. Recent cytochemical and biochemical data indicate that
protein content in peroxisomes are heterogenous and that import of
newly synthesized proteins may be restricted to certain protein
import-competent peroxisomal subcompartments (Yamamoto, K., and Fahimi,
H. D.(1987) J. Cell Biol. 105, 713-722; Heinemann, P.,
and Just, W. W.(1992) FEBS Lett. 300, 179-182;
Lüers, G., Hashimoto, T., Fahimi, H. D., and
Völkl, A.(1993) J. Cell Biol. 121,
1271-1280). We have observed that substantial amounts of
peroxisomal proteins are found together with ``microsomes''
(100,000
Volume 270,
Number 12,
Issue of March 24, 1995 pp. 6949-6958
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
IMPLICATIONS FOR PEROXISOME STRUCTURE AND BIOGENESIS (*)
g pellet) after subcellular fractionation of
rat liver homogenates. In this study we have investigated the origin of
these peroxisomal proteins by modified gradient centrifugation
procedures in Nycodenz and by analysis of enzyme activity
distributions, Western blotting, and immunoelectron microscopy. It is
concluded that much of this material is confined to novel populations
of ``peroxisomes.'' Immunocytochemistry on gradient fractions
showed that some vesicles were enriched in acyl-CoA oxidase and
peroxisomal multifunctional enzyme (``catalase-negative'')
whereas others were enriched in catalase and thiolase (``acyl-CoA
oxidase-negative''). Double immunolabeling experiments verified
the strong heterogeneity in the protein contents of these vesicles and
also identified peroxisomes varying in size from about 0.5 µm
(``normal peroxisomes'') to extremely small vesicles of less
than 100 nm in diameter. The possibility that these vesicles may be
related to different subcompartments of a larger peroxisomal structure
involved in protein import and biogenesis will be discussed.
)
)
)
We thank Dr. Björn Afzelius for
helpful discussions and Dr. Henrik Garoff and Dr. J. Kalervo Hiltunen
for critical reading of this manuscript.
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
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