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Subunit structure of 6-phosphofructokinase from brewers' yeast
Hoppe-Seyler'S Zeitschrift Fur Physiologische Chemie
|November 1, 1975
Summary
Researchers analyzed 6-phosphofructokinase from brewers' yeast using sodium dodecyl sulfate. Hyaluronidase treatment revealed smaller enzyme components, suggesting a complex subunit structure influencing molecular weight determination.
Area of Science:
- Biochemistry
- Enzymology
- Yeast Metabolism
Background:
- 6-phosphofructokinase is a key glycolytic enzyme in yeast.
- Understanding enzyme subunit composition is crucial for elucidating function.
- Sodium dodecyl sulfate (SDS) is commonly used for protein molecular weight determination.
Purpose of the Study:
- To analyze the subunit composition of 6-phosphofructokinase from brewers' yeast.
- To investigate the effect of hyaluronidase treatment on enzyme components.
- To clarify the molecular weight determination of enzyme subunits.
Main Methods:
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
- Ultracentrifugation and gel filtration for molecular weight analysis.
- Enzyme treatment with hyaluronidase.
Main Results:
- Four components (alpha, beta, alpha', beta') with distinct molecular weights were identified.
- Hyaluronidase treatment converted alpha and beta components to alpha' and beta' components.
- SDS-electrophoresis after hyaluronidase treatment yielded the smallest molecular structures, indicating dissociation.
- Hyaluronidase appears to desensitize enzyme components to SDS, affecting apparent molecular weight.
Conclusions:
- The native enzyme likely comprises four subunits (alpha, beta, alpha', beta') or potentially only alpha and beta chains.
- Hyaluronidase treatment is essential for accurate dissociation and molecular weight determination of 6-phosphofructokinase subunits.
- The interaction between enzyme subunits and SDS is modulated by hyaluronidase activity.