An asparaginyl endopeptidase mediates in vivo protein backbone cyclization

Proteases can catalyze both peptide bond cleavage and formation, yet the hydrolysis reaction dominates in nature. This presents an interesting challenge for the biosynthesis of backbone cyclized (circular) proteins, which are encoded as part of precursor proteins and require post-translational peptide bond formation to reach their mature form. The largest family of circular proteins are the plant-produced cyclotides; extremely stable proteins with applications as bioengineering scaffolds. Little is known about the mechanism by which they are cyclized in vivo but a highly conserved Asn (occasionally Asp) residue at the C terminus of the cyclotide domain suggests that an enzyme with specificity for Asn (asparaginyl endopeptidase; AEP) is involved in the process. Nicotiana benthamiana does not endogenously produce circular proteins but when cDNA encoding the precursor of the cyclotide kalata B1 was transiently expressed in the plants they produced the cyclotide, together with linear forms not commonly observed in cyclotide-containing plants. Observation of these species over time showed that in vivo asparaginyl bond hydrolysis is necessary for cyclization. When AEP activity was suppressed, either by decreasing AEP gene expression or using a specific inhibitor, the amount of cyclic cyclotide in the plants was reduced compared with controls and was accompanied by the accumulation of extended linear species. These results suggest that an AEP is responsible for catalyzing both peptide bond cleavage and ligation of cyclotides in a single processing event.

A review of current knowledge of the weedy species Themeda quadrivalvis (grader grass)

This review of grader grass (Themeda quadrivalvis) attempts to collate current knowledge and identify knowledge gaps that may require further research. Grader grass is a tropical annual grass native to India that is now spread throughout many of the tropical regions of the world. In Australia, it has spread rapidly since its introduction in the 1930s and is now naturalised in the tropical areas of Queensland, the Northern Territory and Western Australia and extends south along the east coast to northern New South Wales. It is a vigorous grass with limited palatability, that is capable of invading native and improved pastures, cropping land and protected areas such as state and national parks. Grader grass can form dense monocultures that reduce biodiversity, decrease animal productivity and increase the fire hazard in the seasonally dry tropics. Control options are based on herbicides, grazing management and slashing, while overgrazing appears to favour grader grass. The effect of fire on grader grass is inconclusive and needs to be defined. Little is known about the biology and impacts of grader grass in agricultural and protected ecosystems in Australia. In particular, information is needed on soil seed bank longevity, seed production, germination and growth, which would allow the development of management strategies to control this weedy grass.