What question did this study set out to answer?

To identify why the abundant bone protein osteocalcin and its post-translational modifications evade detection in high-throughput mass spectrometry workflows.

September 5, 2016Rapid Communications in Mass Spectrometry

Influence of carboxylation on osteocalcin detection by mass spectrometry

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To identify why the abundant bone protein osteocalcin and its post-translational modifications evade detection in high-throughput mass spectrometry workflows.
Purified osteocalcin from bovine bone tissue and examined both intact and trypsin-digested forms.
Separated samples using high-performance liquid chromatography and analyzed them via an LTQ-Orbitrap XL mass spectrometer using higher-energy collision dissociation (HCD) fragmentation.
Assigned peptide identities and modification states using Mascot and ProSight Lite software tools.
Confirmed the presence of up to three carboxylation modifications across intact osteocalcin and digested peptide fragments.
Identified osteocalcin peptides using Mascot but failed to pinpoint carboxylation locations due to extensive neutral loss.
Demonstrated via ProSight Lite that intact and fragmented species exhibit complete sequence fragmentation accompanied by total neutral loss of carboxyl moieties.

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Implication

Proteomic analysis reveals that carboxylation-induced neutral loss impairs osteocalcin detection in bovine bone extracts, highlighting the need for specialized mass spectrometry methods.

Cite This Study

Cleland et al. (2016) studied this question.

synapsesocial.com/papers/6a9332ef08fea9c5d8d1866f https://doi.org/10.1002/rcm.7692

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Implication

Proteomic analysis reveals that carboxylation-induced neutral loss impairs osteocalcin detection in bovine bone extracts, highlighting the need for specialized mass spectrometry methods.

Influence of carboxylation on osteocalcin detection by mass spectrometry

Key Points

Discussion

Implication

Cite This Study

Influence of carboxylation on osteocalcin detection by mass spectrometry

Key Points

Discussion

Implication

Cite This Study

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