Isolation and partial characterization of a plasma membrane-enriched fraction of Blastomyces dermatitidis.
The isolation and characterization of a plasma membrane-enriched fraction is reported from the pathogenic fungus Blastomyces dermatitidis employing a simplified and inexpensive procedure. The yeast cells, suspended in an osmotic stabilizer buffer, were subjected to gentle mechanical disruption followed by a three-step differential centrifugation to obtain a 40,000 x g sediment. This fraction showed 2.7- and 3.3-fold relative enrichment of the plasma membrane marker enzymes Mg2(+)-ATPase and 5'-nucleotidase, respectively, with modest cross-contamination of other intracellular membranes. The fraction comprised approximately 49% protein, 31% carbohydrate, 11% phospholipids and 9% sterol with sterol to phospholipid molar ratio of 1.1. Transmission electron microscopy of the plasma membrane-enriched fraction showed homogeneous vesicles with a tripartite unit membrane of 115 A thickness.
Chaturvedi
V P
VP
National Reference Centre for Respiratory Mycoses (ICMR), University of Delhi, India.
Chaturvedi
S
S
Randhawa
H S
HS
Khan
Z U
ZU
Puri
B K
BK
D015720:5'-Nucleotidase / Q000378:metabolism; D001758:Blastomyces / Q000648:ultrastructure*; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism; D002241:Carbohydrates / Q000032:analysis; D002458:Cell Fractionation; D002462:Cell Membrane / Q000032:analysis / Q000201:enzymology / Q000648:ultrastructure*; D008563:Membrane Lipids / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D008854:Microscopy, Electron; D010743:Phospholipids / Q000032:analysis
D002241:Carbohydrates; D008563:Membrane Lipids; D008565:Membrane Proteins; D010743:Phospholipids; D015720:5'-Nucleotidase; D017301:Ca(2+) Mg(2+)-ATPase