The importance of protein-protein interactions for optimising oxygen activity in photosystem II: reconstitution with a recombinant thioredoxin--manganese stabilising protein.

In this paper we describe how photosystem II (PSII) from higher plants, which have been depleted, of the extrinsic proteins can be reconstituted with a chimeric fusion protein comprising thioredoxin from Escherichia coli and the manganese stabilising protein from Thermosynechococcus elongatus. Surprisingly, even though E. coli thioredoxin is completely unrelated to PSII, the fusion protein restores higher rates of activity upon rebinding to PSII than either the native spinach MSP, or T. elongatus MSP. PSII reconstituted with the fusion protein also has a lower requirement for calcium than PSII with the small extrinsic proteins removed, or PSII reconstituted with spinach or T. elongatus MSP. The MSP portion of the fusion protein is less thermally stable compared to isolated MSP from T. elongatus, which could be the key to its superior activation capability through greater flexibility. This work reveals the importance of protein-protein interactions in the water splitting activity of PSII and suggests that conformational configurations, which increase flexibility in MSP, are essential to its function, even when these are induced by an unrelated protein.

Growth responses of cartilage to static and dynamic compression.

During skeletal development, growth, and maturation, gradual changes in the material properties and physical dimensions of cartilage occur under the influence of mechanical loading. The objective of the current study was to compare glycosaminoglyean biosynthesis and cell proliferation in fetal, calf, and adult bovine cartilage explants, isolated from defined depths from the articular surface, in response to controlled compressive loads. Mechanical testing confirmed that for all cartilage samples subjected to load, there was a marked time-averaged (static) compression, whereas the addition of dynamic load at a frequency of 0.01 Hz induced dynamic strain with amplitude and phase shift characteristics typical of stimuli that previously were found to be associated with stimulation of glycosaminoglycan synthesis. In metabolic studies, the application of static loading (84 kPa) for 24 hours inhibited glycosaminoglycan and deoxyribonucleic acid synthesis in all cultured cartilage samples. The superposition of dynamic loading (200 kPa, 0.01 Hz) induced a 20% stimulation of glycosaminoglycan biosynthesis in calf cartilage from the middle-deep zones over statically-loaded samples and an additional approximate 50% suppression of deoxyribonucleic acid synthesis in fetal and calf cartilage from the articular surface. These results indicate that synthesis of glycosaminoglycan and deoxyribonucleic acid, two distinct indices of cartilage growth, are regulated independently by mechanical loading and that cartilage responds differently to static and dynamic loading at different stages of maturation.

Compressive properties and function-composition relationships of developing bovine articular cartilage.

The composition of cartilage is known to change during fetal and postnatal development. The objectives of this study were to characterize the compressive biomechanical properties of the 1 mm thick articular layer of cartilage of the distal femur from third-trimester bovine fetuses, from 1 to 3 week old bovine calf and from young adult bovine knees, and to correlate these properties with tissue components. The confined compression modulus increased 180% from the fetus to the calf and adult. The hydraulic permeability at 45% offset compression (relative to the free-swelling thickness) decreased by 70% from fetus to adult. These development-associated changes in biomechanical properties were primarily associated with a marked (approximately 2-3-fold) increase during development in collagen content and no detectable change in glycosaminoglycan (GAG) content. A role for collagen in the compressive properties of cartilage and the gradual increase in collagen during development suggest that collagen metabolism is critical for cartilage tissue engineering and repair therapies.