Coexpression of connexin 45 with connexin 43 decreases gap junction size.

In the human heart, ventricular myocytes express connexin 43 (Cx43) and traces of Cx45. In congestive heart failure, Cx43 levels decrease, Cx45 levels increase and gap junction size decreases. To determine whether alterations of connexin coexpression ratio influence gap junction size, we engineered a rat liver epithelial cell line that endogenously expresses Cx43 to coexpress inducible levels of Cx45 under stimulation of the insect hormone, ponasterone A.

Remodelling of gap junctions and connexin expression in diseased myocardium.

Gap junctions form the cell-to-cell pathways for propagation of the precisely orchestrated patterns of current flow that govern the regular rhythm of the healthy heart. As in most tissues and organs, multiple connexin types are expressed in the heart: connexin43 (Cx43), Cx40 and Cx45 are found in distinctive combinations and relative quantities in different, functionally-specialized subsets of cardiac myocyte. Mutations in genes that encode connexins have only rarely been identified as being a cause of human cardiac disease, but remodelling of connexin expression and gap junction organization are well documented in acquired adult heart disease, notably ischaemic heart disease and heart failure.

Gap junction remodelling in human heart failure is associated with increased interaction of connexin43 with ZO-1.

Aims: Remodelling of gap junctions, involving reduction of total gap junction quantity and down-regulation of connexin43 (Cx43), contributes to the arrhythmic substrate in congestive heart failure. However, little is known of the underlying mechanisms. Recent studies from in vitro systems suggest that the connexin-interacting protein zonula occludens-1 (ZO-1) is a potential mediator of gap junction remodelling.

Functional characterisation of a nematode secreted GM2-activator protein.

We have identified a GM2-activator protein (GM2AP) with highly unusual properties secreted by the nematode parasite Trichinella spiralis. Expression in Pichia pastoris resulted in a hyperglycosylated protein of 28 kDa, but the 18 kDa native protein was not glycosylated. The parasite GM2AP does not facilitate degradation of GM2 ganglioside by N-acetyl-beta-hexosaminidase A, although it does inhibit phospholipase D activity.

Characterisation of a secreted N-acetyl-beta-hexosaminidase from Trichinella spiralis.

A thorough investigation was conducted for glycoside hydrolase activities in the secreted proteins of Trichinella spiralis. The data demonstrated that the only secreted glycosidase with significant activity was an exo-beta-hexosaminidase with catalysis of the substrates N-acetyl-beta-D-glucosamine, N-acetyl-beta-D-galactosamine and N-acetyl-beta-D-glucosamine-6-sulphate proceeding with an efficiency similar to the human isozyme beta-hexosaminidase A (Hex A). The hydrolysis of N-acetyl-beta-D-glucosamine followed Michaelis-Menten kinetics with a K(m) of 0.