A common beta-sheet architecture underlies in vitro and in vivo beta(2)-microglobulin amyloid fibrils

Misfolding and aggregation of normally soluble proteins into amyloid fibrils and their deposition and accumulation underlies a variety of clinically significant diseases. Fibrillar aggregates with amyloid-like properties can also be generated in vitro from pure proteins and peptides, including those not known to be associated with amyloidosis. Whereas biophysical studies of amyloid-like fibrils formed in vitro have provided important insights into the molecular mechanisms of amyloid generation and the structural properties of the fibrils formed, amyloidogenic proteins are typically exposed to mild or more extreme denaturing conditions to induce rapid fibril formation in vitro. Whether the structure of the resulting assemblies is representative of their natural in vivo counterparts, thus, remains a fundamental unresolved issue. Here we show using Fourier transform infrared spectroscopy that amyloid-like fibrils formed in vitro from natively folded or unfolded β2-microglobulin (the protein associated with dialysis-related amyloidosis) adopt an identical β-sheet architecture. The same β-strand signature is observed whether fibril formation in vitro occurs spontaneously or from seeded reactions. Comparison of these spectra with those of amyloid fibrils extracted from patients with dialysis-related amyloidosis revealed an identical amide I' absorbance maximum, suggestive of a characteristic and conserved amyloid fold. Our results endorse the relevance of biophysical studies for the investigation of the molecular mechanisms of β2-microglobulin fibrillogenesis, knowledge about which may inform understanding of the pathobiology of this protein

Sustainable Harvesting of Tropical Rainforests: Reply to Keto, Scott and Olsen

This paper refutes the Keto et al. proposition that the Queensland selection logging system is neither ecologically nor economically sustainable. The key requirements of this system are: (1) that logging guidelines are sympathetic to the silvicultural characteristics of the forest, ensuring adequate regeneration of commercial species and discouraging invasion by weeds; (2) tree-marking by trained staff specifies trees to be retained, trees to be removed and the direction of felling to ensure minimal damage to the residual stand; (3) logging equipment is appropriate and driven by trained operators to ensure minimal damage and soil disturbance, compaction and erosion; (4) prescriptions ensure that adequate stream buffers and steep slopes are excluded from logging; (5) sufficient areas for scientific reference, feature protection and recreation are identified and excluded from logging; and (6) that deficiencies in an evolving system are recognized and remedied, leading to an improved system. Many studies of the effects of logging in these forests have been published and collectively provide a unique demonstration of one possible approach to sustainable timber harvesting

Structural and functional aspects of the pentraxins and SAA in the*acute-phase response and amyloidosis

SAA - serum amyloid A proteinSIGLEAvailable from British Library Document Supply Centre-DSC:DXN006717 / BLDSC - British Library Document Supply CentreGBUnited Kingdo

A Systematic study of the effect of physiological factors on beta2-microglobulin amyloid formation at neutral pH

Noß2-microglobulin (ß2m) forms amyloid fibrils that deposit in the musculo-skeletal system in patients undergoing long-term hemodialysis. How ß2m self-assembles in vivo is not understood, since the monomeric wild-type protein is incapable of forming fibrils in isolation in vitro at neutral pH, while elongation of fibril-seeds made from recombinant protein has only been achieved at low pH or at neutral pH in the presence of detergents or cosolvents. Here we describe a systematic study of the effect of 11 physiologically relevant factors on ß2m fibrillogenesis at pH 7.0 without denaturants. By comparing the results obtained for the wild-type protein with those of two variants (¿N6 and V37A), the role of protein stability in fibrillogenesis is explored. We show that ¿N6 forms low yields of amyloid-like fibrils at pH 7.0 in the absence of seeds, suggesting that this species could initiate fibrillogenesis in vivo. By contrast, high yields of amyloid-like fibrils are observed for all proteins when assembly is seeded with fibril-seeds formed from recombinant protein at pH 2.5 stabilized by the addition of heparin, serum amyloid P component (SAP), apolipoprotein E (apoE), uremic serum, or synovial fluid. The results suggest that the conditions within the synovium facilitate fibrillogenesis of ß2m and show that different physiological factors may act synergistically to promote fibril formation. By comparing the behavior of wild-type ß2m with that of ¿N6 and V37A, we show that the physiologically relevant factors enhance fibrillogenesis by stabilizing fibril-seeds, thereby allowing fibril extension by rare assembly competent species formed by local unfolding of native monomers