Journal of Bioenergetics and Biomembranes

, Volume 26, Issue 4, pp 447–456 | Cite as

Preparation of highly phosphorylating mitochondria from the yeastSchizosaccharomyces pombe

  • Jean-Michel Jault
  • Jane Comte
  • Danièle C. Gautheron
  • Attilio Di Pietro
Original Articles
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Abstract

Schizosaccharomyces pombe yeast cells grown on either fermentable or respiratory media were efficiently converted to stable spheroplasts by the α-(1 → 3)-glucanase Novozym 234 in the presence of 1.2M sorbitol. Lysis of spheroplasts by gentle homogenization in dilute sorbitol resulted in the preparation of mitochondria with a structure similar to that observed within the starting yeast cells. The isolated mitochondria exhibited high oxidation rates with various respiratory substrates, NADH being the most efficient. The mitochondria appeared well coupled since the second State 4 rate observed after ADP consumption was identical to the initial one. The State 3 rate in the presence of ADP was completely inhibited by low oligomycin concentrations, similarly to the concomitant ATP synthesis of 900 nmol/min × mg protein. These NADH oxidation and dependent ATP-synthesis activities are much higher than those previously described for mitochondria isolated fromSchizosaccharomyces pombe, and similar to the highest values reported forSaccharomyces cerevisiae.

Key words

Isolated mitochondria yeastSchizosaccharomyces pombe spheroplasts oxidative phosphorylation respiration ATP synthesis 
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Copyright information

© Plenum Publishing Corporation 1994

Authors and Affiliations

  • Jean-Michel Jault
    • 1
  • Jane Comte
    • 2
  • Danièle C. Gautheron
    • 1
  • Attilio Di Pietro
    • 2
  1. 1.Campus de La Doua, Laboratoire de Biologie et Technologie des Membranes et des Systèmes Intégrés, VilleurbanneUniversité Claude Bernard-Lyon IFrance
  2. 2.Campus de Gerland, Institut de Biologie et Chimie des Protéines, LyonUniversité Claude Bernard-Lyon IFrance

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