Probing short and long-range interactions in native collagen inside the bone matrix by BioSolids CryoProbe

Area of Science:

• Biomaterials Science
• Biophysics
• Structural Biology

Background:

• Solid-state nuclear magnetic resonance (NMR) is crucial for investigating bone mineralization and collagen interactions.
• Low sensitivity in conventional NMR probes limits detailed structural analysis of native collagen.
• Advancements are needed to overcome sensitivity limitations in solid-state NMR for biomaterials.

Purpose of the Study:

• To evaluate the applicability of a newly developed BioSolids CryoProbe™ for solid-state NMR.
• To elucidate atomic-level structural details of native collagen within bone.
• To assess sensitivity enhancements offered by the cryoprobe for biomaterial analysis.

Main Methods:

• Solid-state NMR experiments utilizing the BioSolids CryoProbe™.
• Measurement of natural abundance ¹³C spectrum sensitivity.
• Performance of natural abundance ¹⁵N cross-polarization magic angle spinning (CPMAS) experiments.
• Execution of 2D ¹H-¹³C heteronuclear correlation (HETCOR) experiments on native collagen.

Main Results:

• Approximately a fourfold sensitivity enhancement was achieved in the ¹³C spectrum compared to conventional probes.
• The enhanced sensitivity enabled natural abundance ¹⁵N CPMAS and 2D ¹H-¹³C HETCOR experiments within a reasonable timeframe.
• 2D ¹H/¹³C HETCOR experiments successfully detected numerous short- and long-range interactions in native collagen assembly.

Conclusions:

• The BioSolids CryoProbe™ significantly improves sensitivity in solid-state NMR for biomaterials.
• This enhanced sensitivity facilitates detailed structural investigations of native collagen.
• The method's scope is expanded for analyzing challenging biomaterials like bone collagen.