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Updated: Aug 21, 2026

Actin Co-Sedimentation Assay; for the Analysis of Protein Binding to F-Actin
Published on: March 27, 2008
Characterization of boar sperm cytoskeletal cylicin II as an actin-binding protein
Roselyne Rousseaux-Prévost1, Christophe Lécuyer, Hervé Drobecq
1FRC3 CNRS, Institut de Biologie de Lille-BP 447, Lille 59021, France. roselyne.rousseaux@ibl.fr
Abstract:
The presence of actin-binding proteins in the perinuclear theca of boar spermatozoa has been investigated, using stepwise extractions of proteins from sperm heads. Proteins extracted with the alkaline buffer 1M Na(2)CO(3), pH 11, were found to contain a 66kDa protein that binds F-actin in actin pelleting assays. Sequence studies and immunological characterization with antibodies specific for human cylicin II identified the 66kDa protein as the homologue of bovine and human cylicin II. Immunocytochemical studies showed the presence of porcine cylicin II in the acrosomal region of round spermatids and in the postacrosomal region of late spermatids and spermatozoa, in agreement with the previously described localization of cylicins. Taken together, the results suggest that cylicin II, a protein of the sperm perinuclear cytoskeleton, is a novel actin-binding protein, which probably plays a role in the actin-related events that occur during spermiogenesis and the early events of fertilization.
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