Characterization of proADAMTS5 processing by proprotein convertases.

ADAMTS5 (aggrecanase-2), a key metalloprotease mediating cartilage destruction in arthritis, is synthesized as a zymogen, proADAMTS5. We report a detailed characterization of the propeptide excision mechanism and demonstrate that it is a major regulatory step with unusual characteristics. Using furin-deficient cells and a furin inhibitor, we found that proADAMTS5 was processed by proprotein convertases, specifically furin and PC7, but not PC6B. Mutagenesis of three sites containing basic residues within the ADAMTS5 propeptide (RRR⁴⁶, RRR⁶⁹ and RRRRR²⁶¹) suggested that proADAMTS5 processing occurs after ^Arg261. That furin processing was essential for ADAMTS5 activity was illustrated using the known ADAMTS5 substrate aggrecan, as well as a new substrate, versican, an important regulatory proteoglycan during mammalian development. When compared to other ADAMTS proteases, proADAMTS5 processing has several distinct features. In contrast to ADAMTS1, whose furin processing products were clearly present intracellularly, cleaved ADAMTS5 propeptide and mature ADAMTS5 were found exclusively in the conditioned medium. Despite attempts to enhance detection of intracellular proADAMTS5 processing, such as by immunoprecipitation of total ADAMTS5, overexpression of furin, and secretion blockade by monensin, neither processed ADAMTS5 propeptide nor the mature enzyme were found intracellularly, which was strongly suggestive of extracellular processing. Extracellular ADAMTS5 processing was further supported by activation of proADAMTS5 added exogenously to HEK293 cells stably expressing furin. Unlike proADAMTS9, which is processed by furin at the cell-surface, to which it is bound, ADAMTS5 does not bind the cell-surface. Thus, the propeptide processing mechanism of ADAMTS5 has several points of distinction from those of other ADAMTS proteases, which may have considerable significance in the context of osteoarthritis.

A critical review of relations between corporate responsibility research and practice

This essay identifies epistemological, theoretical and methodological problems in a potentially influential subset of the interdisciplinary corporate responsibility literature, that which appears in the management literature. The received conceptualization of stakeholder analysis is criticised by identifying six sets of factors conventionally considered as promoting social responsibilities in the firm: inter-organizational factors, economic competitors, institutional investors, end-consumers, government regulators and non-governmental organizations. Each is addressed on conceptual grounds, its empirical salience in terms of the latest relevant research and prospects to be a significant factor in promoting outcomes consistent with social welfare. Despite obvious antagonistic relations between organization-centred economic objectives and extra-organizational-directed social considerations, the huge body of research we address drifts in a disengaged Sargasso Sea. The essay argues for appropriate directions for continuing business ethics/responsibility/corporate citizenship research, suggesting certain sociological works on moral leadership, moral courage, and academic leadership.

A method and its optimal parameters for the measurement of mass transfer coefficient in heat treatment furnaces

The mass transfer coefficient is an important kinetic factor to control the thermo-chemical treatment processes of metals and alloys. More importantly, the mass transfer coefficient is different at different surface positions of a metallic part treated, which depends on the dynamic characteristics of the atmosphere close to the treated surface. Understanding the local mass transfer coefficient would be significant to approach the expected physical and mechanical properties of treated surfaces. In this paper, a reverse method was proposed to measure the mass transfer coefficient at component surface and the diffusivity in metal during heat treatment. The methodology of the reverses method and the optimal parame-ters are discussed in some detail. This method was successfully used to determine the car-bon transfer coefficient at the surface of a part in a carburizing furnace and carbon diffusiv-ity from the carbon distribution within the diffusion layer.