Kinetics of thermal inactivation of catalase in the presence of additives

The kinetics of catalase inactivation in buffer-free aqueous solutions within the temperature range 30–60 °C in the absence or presence of hydrogen peroxide was investigated and discussed taking into account the effect of NaCl, ethane-1,2-diol, propane-1,2,3-triol and sucrose, additives expected to increase the enzyme thermostability. Using the kinetic extended curves obtained by measuring the substrate absorbance in time and an isoconversional method, several simple kinetic models most frequently encountered in literature were fitted to the experimental data. The best model for inactivation was chosen on the basis of several statistical criteria. The half-times of inactivation and the activation energies were also reported and discussed.     

Mutational and structural analyses of the hinge region of membrane type 1-matrix metalloproteinase and enzyme processing

Membrane type 1 (MT1)-matrix metalloproteinase (MMP) is a major mediator of collagen degradation in the pericellular space in both physiological and pathological conditions. Previous evidence has shown that on the cell surface, active MT1-MMP undergoes autocatalytic processing to a major membrane-tethered 44-kDa product lacking the catalytic domain and displaying Gly285 at its N terminus, which is at the beginning of the hinge domain. However, the importance of this site and the hinge region in MT1-MMP processing is unknown. In the current study, we generated mutations and deletions in the hinge of MT1-MMP and followed their effect on processing. These studies established Gly284-Gly285 as the main cleavage site involved in the formation of the 44-kDa species. However, alterations at this site did not prevent processing. Instead, they forced downstream cleavages within the stretch of residues flanked by Gln296 and Ser304 in the hinge region, as determined by the processing profile of various hinge deletion mutants. Also, replacement of the hinge of MT1-MMP with the longer MT3-MMP hinge did not prevent processing of MT1-MMP. Molecular dynamic studies using a computational model of MT1-MMP revealed that the hinge region is a highly motile element that undergoes significant motion in the highly exposed loop formed by Pro295-Arg302 consistent with being a prime target for proteolysis, in agreement with the mutational data. These studies suggest that the hinge of MT1-MMP evolved to facilitate processing, a promiscuous but compulsory event in the destiny of MT1-MMP, which may play a key role in the control of pericellular proteolysis.     

The role of connexins in controlling cell growth and gene expression

The purpose of this paper is to provide a brief overview of current thinking on the role of connexins, in particular Cx43, in growth regulation, and a more detailed discussion as to potential mechanisms involved with an emphasis on gene expression. While the precise molecular mechanism by which connexins can affect the growth of normal or tumor cells remains elusive, a number of exciting reports have expanded our understanding and are presented in some detail. Thus, we will discuss (Section 2): the role of protein-protein interactions in integrating connexins into multiple signal transduction pathways; phosphorylation at specific sites and reversal of growth inhibition; the role of the carboxy-terminal regulatory domain as a signaling molecule. Some of our latest work on the potential functions of endogenously produced carboxy-terminal fragments of Cx43 are also presented (Section 3). Finally, Section 4 will pay tribute to the rapidly emerging realization that connexins such as Cx43 and Cx32 exert important and extensive effects on gene expression, particularly those genes linked to growth regulation.     

Infections with human rhinovirus induce the formation of distinct functional membrane domains.

The plasma membrane contains distinct domains that are characterized by a high concentration of sphingolipids and cholesterol. These membrane microdomains also referred to as rafts, seem to be intimately involved in transmembranous signaling and often initiate interactions of pathogens and the host cell membranes. Here, we investigated the further reorganization of membrane rafts in cultured epithelial cells and ex vivo isolated nasal cells after infection with rhinoviruses. We demonstrate the formation of ceramide-enriched membrane platforms and large glycosphingolipid-enriched membrane domains and the co-localization of fluorochrome-labeled rhinoviruses with these membrane domains during attachment and uptake of human rhinovirus. Destruction of glycosphingolipid-enriched membrane domains blocked infection of human cells with rhinovirus. Furthermore, our studies indicate that the activation of the acid sphingomyelinase (ASM) is intrigued in the formation of ceramide- or GM1- enriched membrane platforms. Inhibition of the ASM reduces the number of ceramide-enriched platforms and glycosphingolipid-enriched membrane domains. These data reveal a critical role of the ASM for the formation of membrane platforms and infection of human cells with rhinoviruses.     

Oxidase–peroxidase reaction: kinetics of peroxidase-catalysed oxidation of 2-aminophenol

Possible reaction pathways that may lead to horseradish peroxidase inactivation during the aerobic oxidation of 2-aminophenol were investigated using extended kinetic curves. A kinetic model involving the formation of a low-reactive species, Compound III, was proposed and several rate constants were calculated using an optimisation computing program. Sensitivity analysis allowed to conclude that both oxidase and peroxidase cycles occur in 2-aminophenol oxidation.     

Halophilic archaea isolated from man-made young (200 years) salt lakes in Slănic, Prahova, Romania

We isolated a number of extremely halophilic archaea from four salt lakes (Red Bath, Green Bath, Shepherd Bath and Bride Cave) located in Slanic Prahova, Romania. The characterization of 12 representative isolates by polyphasic approach revealed that 11 strains were members of the genus Haloferax and only one was a member of the genus Haloarcula. The 11 Haloferax isolates possessed sulfated diglycosylarchaeol-1 as the major membrane glycolipid, and G+C contents of total DNA were 63.4–65.8 mol%. The predominant isolation of Haloferax species from the lakes may suggest that the underground salt deposit possesses Haloferax species as the major biota of ancient origin. To the best of the authors’ knowledge, this is the first paper on the survey of halophilic archaea of man-made young salt lakes.     

Insulin resistance and decreased spexin in Indian Patients with Type 2 Diabetes Mellitus

Spexin is novel biomarker, which plays a potential role in glucose and lipid metabolisms. However, there was paucity of serum spexin levels in obesity and diabetes mellitus subjects. Hence the current study was aimed to find the relationship between the serum spexin levels in type 2 Diabetes mellitus (type 2 DM) with extrapolation of cardiovascular disease (CVD) risk. A cross-sectional study included 330 participants, subdivided as control (n=110), type 2 DM (n=110) and type 2 DM with CVD groups (n=110). HbA1c, insulin, lipid profile, spexin & leptin including blood pressure and body mass index were analyzed from all the participants. The serum spexin levels (ng/ml) were significantly decreased in type 2 DM (mean ± sd: 0.65 ± 0.03) and type 2 DM with CVD (0.48 ± 0.02) groups compared to the control (0.79 ± 0.03) group (p<0.001). The decreased spexin levels were observed in type 2 DM, and further more decreased in type 2 DM with CVD patients compared to controls indicating that spexin levels could be served as an early prediction of obesity-induced T2DM with CVD risk.