Purification of guinea pig small intestinal peroxisomes and the subcellular localization of glucose-6-phosphate dehydrogenase.
A method for the isolation of highly purified peroxisomes from guinea pig small intestine was developed. This two-stage process involved a rate-dependent banding of a light-mitochondria lambda-fraction followed by a density-dependent banding of the catalase enriched fractions obtained from the first step, using a horizontal rotor. Furthermore, the subcellular localization of glucose-6-phosphate dehydrogenase (NADP+-dependent) activity in guinea pig small intestine was examined. Analysis of density-gradient fractions indicated that approximately 3-4% of the cellular NADP+-dependent glucose-6-phosphate dehydrogenase activity is associated with peroxisomal fractions and that it is localized to the matrix of peroxisomes. It is therefore suggested that a peroxisomal source of NADPH may be utilized by enzyme systems that use NADPH specifically as a reductant.
Tappia
P S
PS
Institute of Cardiovascular Sciences, Department of Human Anatomy, University of Manitoba, Winnipeg, Canada.
Jones
C J
CJ
Connock
M J
MJ
D000818:Animals; D015415:Biomarkers / Q000737:chemistry; D002458:Cell Fractionation / Q000379:methods; D002499:Centrifugation, Density Gradient; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism*; D006168:Guinea Pigs; D007421:Intestine, Small / Q000378:metabolism*; D008297:Male; D008793:Metrizamide / Q000378:metabolism; D008830:Microbodies / Q000201:enzymology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D009249:NADP / Q000378:metabolism; D017830:Octoxynol / Q000494:pharmacology
D015415:Biomarkers; D009249:NADP; D017830:Octoxynol; D005954:Glucosephosphate Dehydrogenase; D008793:Metrizamide