In guinea pig sperm, aldolase A forms a complex with actin, WAS, and Arp2/3 that plays a role in actin polymerization

Abstract

Glycolytic enzymes have, in addition to their role in energy production, other functions in the regulation of cellular processes. Aldolase A has been reported to be present in sperm, playing a key role in glycolysis; however, despite its reported interactions with actin and WAS, little is known about a non-glycolytic role of aldolase A in sperm. Here, we show that in guinea pig spermatozoa, aldolase A is tightly associated to cytoskeletal structures where it interacts with actin, WAS, and Arp2/3. We show that aldolase A spermatozoa treatment increases their polymerized actin levels. In addition, we show that there is a direct correlation between the levels of polymerized actin and the levels of aldolase A-actin interaction. Our results suggest that aldolase A functions as a bridge between filaments of actin and the actin-polymerizing machinery.

Metabolic Optimization by Enzyme-Enzyme and Enzyme-Cytoskeleton Associations

“…Such an effect has been reported for aldolase (Chiquete-Felix et al, 2009;Schindler et al, 2001). An interesting finding is that some growth factors, such as PGF and EGF enhance the GAPDH/cytoskeleton interaction, possibly increasing keratinocyte migration (Tochio et al, 2010).…”

Wolbachia pipientis grows in Saccharomyces cerevisiae evoking early death of the host and deregulation of mitochondrial metabolism

“…Samples were diluted in a 4X buffer (500 mmol L −1 Tris, pH 6.8, 10% glycerol, 10% SDS, 0.05% beta‐mercapto‐ethanol, and 0.01% bromophenol blue) and boiled for 5 min. SDS/PAGE was performed in 10% polyacrylamide gels and electrotransferred to poly(vinylidenedifluoride) membranes as reported elsewhere (Chiquete‐Felix et al., ). Membranes were blocked with 5% Blotto nonfat dry milk in TBS‐T (50 mmol L −1 Tris, 104 mmol L −1 NaCl, pH 7.6, 0.1% Tween 20) for 1 hr, and incubated overnight at 4°C with the primary antibody.…”

“…Membranes were washed with TBS‐T and incubated at 37°C for 1 hr with secondary antibody. Membranes were washed again and the bands were developed by chemiluminescence using an ECL kit (Amersham Biosciences, GE, Healthcare) (Chiquete‐Felix et al., ). PVDF membranes were stripped as indicated by abcam protocol using a mild‐stripping buffer, blocked with 5% Blotto nonfat dry milk in TBS‐T and reprobed with a different antibody as indicated.…”

Role of Actin Cytoskeleton During Mammalian Sperm Acrosomal Exocytosis

“…In guinea pig sperm, it was claimed that aldolase A is tightly associated with cytoskeletal structures where it interacts with actin and Arp2/3 (Chiquete-Felix et al 2009), suggesting that aldolase A functions as a bridge between filaments of actin and the actin-polymerizing machinery.…”