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Actin-like protein associated with plasma membranes fromEuglena gracilis

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Summary

Microtubules are characteristic components of the membrane skeleton ofEuglena gracilis, but whether microfilaments are present has been controversial. We here present evidence that an actin-like protein may indeed be associated with the plasma membrane (PM) ofE. gracilis. Firstly, a 47 kDa, PM-associated, polypeptide was recognized by an anti-amoeba actin antibody. Secondly, this 47 kDa protein seemed to be peripherally attached to PM in much the same way as β-tubulin, since both could be released from PM by treatment with 150 mM NaOH but not with ethylene glycol, NaCl, or formamide. Thirdly, the 47 kDa polypeptide and β-tubulin were found mainly in the Triton X-1 14-insoluble fraction, indicating that they were part of a protein complex resistant to detergents, such as the cytoskeleton. Finally, DNase I activity was inhibited by a fraction enriched in the 47 kDa polypeptide, a property typical of actin.

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Abbreviations

CP-medium:

cytoskeleton preparation medium

BNSP-skatole:

2-(2′-nitrophenylsulfenyl)-3-methyl-3′-bromoindolenine

ECL:

enhanced chemiluminescence

HEPES:

N-[2-hydroxyethyl]-piperazine-N′[2-ethane sulfonic acid]

ICM:

intracellular membranes

MF:

mitochondrial and microsomal fraction

PM:

plasma membrane

PPB:

potassium phosphate buffer

PVDF:

polyvinylidene difluoride

SDS:

sodium dodecyl sulphate

TBS:

Tris-buffered saline

TBST:

Tris-buffered saline with Tween 20

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Authors and Affiliations

  1. Section of Plant Physiology, Department of Plant Biology, Lund University, P.O. Box 117, S-221 00, Lund, Sweden

    Silja K. Petersen-Mahrt, Anders Sonesson & Susanne Widell

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  1. Silja K. Petersen-Mahrt
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  2. Anders Sonesson
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  3. Susanne Widell
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Petersen-Mahrt, S.K., Sonesson, A. & Widell, S. Actin-like protein associated with plasma membranes fromEuglena gracilis . Protoplasma 202, 153–160 (1998). https://doi.org/10.1007/BF01282543

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  • DOI: https://doi.org/10.1007/BF01282543

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