ISSN:
1432-072X
Keywords:
Hansenula polymorpha
;
Peroxisomes
;
Methanol
;
Dihydroxyacetone synthase
;
Cell fractionation
;
Immunocytochemistry
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract The subcellular localization of dihydroxyacetone synthase (DHAS) in the methylotrophic yeast Hansenula polymorpha was studied by various biochemical and immunocytochemical methods. After cell fractionation involving differential and sucrose gradient centrifugation of protoplast homogenates prepared from methanol-grown cells, DHAS cosedimented with the peroxisomal enzymes alcohol oxidase and catalase. Electron microscopy of this fraction showed that it contained mainly intact peroxisomes, whereas SDS-polyacrylamide gel electrophoresis revealed two major protein bands (75 and 78 kDa) which were identified as alcohol oxidase and DHAS, respectively. The localization of DHAS in peroxisomes was further established by immunocytochemistry. After immuno-gold staining carried out on ultrathin sections of methanol-grown H. polymorpha using DHAS-specific antibodies, labelling was confined to the peroxisomal matrix.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00411242
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