Enzymatic colicins such as colicin E9 (ColE9) bind to BtuB on the cell surface of Escherichia coli and rapidly recruit a second coreceptor, either OmpF or OmpC, through which the N-terminal natively disordered region (NDR) of their translocation domain gains entry into the cell periplasm and interacts with TolB. Previously, we constructed an inactive disulfide-locked mutant ColE9 (ColE9(s-s)) t...

Tissue functions and mechanical coupling of cells must be integrated throughout development. A striking example of this coupling is the interactions of body wall muscle and hypodermal cells in Caenorhabditis elegans. These tissues are intimately associated in development and their interactions generate structures that provide a continuous mechanical link to transmit muscle forces across the hyp...

BACKGROUND The high level of excretion and rapid folding ability of β-fructofuranosidase (β-FFase) in Escherichia coli has suggested that β-FFase from Arthrobacter arilaitensis NJEM01 can be developed as a fusion partner. METHODS Based on the modified Wilkinson and Harrison algorithm and the preliminary verification of the solubility-enhancing ability of β-FFase truncations, three β-FFase tru...

The TupABC system is involved in the cellular uptake of tungsten and belongs to the ABC (ATP binding cassette)-type transporter systems. The TupA component is a periplasmic protein that binds tungstate anions, which are then transported through the membrane by the TupB component using ATP hydrolysis as the energy source (the reaction catalyzed by the ModC component). We report the heterologous ...

A fusion protein of human interleukin-2 (hIL-2) and green fluorescent protein (GFP) was expressed in insect Sf-9 cells infected with recombinant baculovirus derived from the Autographa californica nuclear polyhedrosis virus (AcNPV). This fusion protein was comprised of a histidine affinity ligand for simplified purification using immobilized metal affinity chromatography (IMAC), UV-optimized GF...

The fundamental mechanism of disease in pancreatitis is autodigestion of pancreatic tissue by digestive enzymes that are prematurely activated within the pancreatic acinar cells. Normally, pancreatic digestive enzymes are released from the pancreas as inactive precursor zymogen granules. Trypsinogen within these granules is proteolytically cleaved and activated when it reaches the duodenal lume...

OBJECTIVE To study the therapeutic application of guanylate cyclase inhibition by methylene blue in an experimental model of acute pancreatitis in pigs. METHODS acute necrotizing pancreatitis was induced in anesthetized pigs by the retrograde infusion of 1 ml/kg of 5% sodium taurocholate and 8 U/kg enterokinase in the pancreatic duct. Three groups were studied (n = 5): control (C), pancreatit...

Thrombokinase of the blood, while resembling enterokinase in its role of activator, is more closely analogous to trypsin in its intrinsic origin. It probably arises from a plasma precursor; but it is different from plasmin (fibrinolysin). Like trypsin, thrombokinase can activate prothrombin without the aid of other factors; however, it is potentiated by platelets plus calcium. Unlike certain ti...

Purification of recombinant proteins has been greatly simplified in recent years because of the availability of expression vectors that allow fusion of the protein coding sequence of interest to short peptide sequences or larger proteins, enabling the affinity purification of the fusion protein from crude preparations. Fusion proteins are often engineered to contain short linkers of five to six...

CONTEXT All work on human trypsinogen activation peptide (TAP) in acute pancreatitis has been carried out with the same assay. Despite the extensive use of this original TAP assay, there is no characterization of the TAP-like immunoreactivity measured. OBJECTIVE The aims of this study were to develop an additional TAP assay and to attempt to characterize the TAP-like immunoreactivity found in...