Unfolding Mechanics of Multiple OspA Substructures Investigated with Single Molecule Force Spectroscopy
作者： Rukman Hertadi , Franz Gruswitz , Lin Silver , Akiko Koide , Shohei Koide
DOI: 10.1016/J.JMB.2003.09.010
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摘要: Abstract We investigated mechanical unfolding of Borrelia burgdorferi outer surface protein A (OspA), a Lyme disease antigen containing unique single-layer β-sheet, with atomic force microscopy (AFM). mechanically stretched monomeric unit, rather than tandem repeat, by pulling it from its N and C-terminal residues without using intervening polymer as spacer. detected two peaks in the force-extension profile before final rupture fully extended polypeptide, which we interpreted multiple substructures OspA. The double-peaked curves are consistent results previous thermodynamic studies showing cooperative units processes were reversible, refolded within one second. Mutations near boundary reduced height first peak to undetectable levels marginally affected second one, indicating that between is related previously assigned units. Based on “worm-like chain” analysis our AFM data, propose model for OspA, where nearly half chain minimal resistive force, followed sequential breakdown N-terminal substructures. these results, discuss similarities differences reactions This work demonstrates study proteins can elucidate details intramolecular mechanics
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