Dataset Information

A Novel Method for Isolating Whole Protein from Human Cranial Bone

ABSTRACT: The presence of the extracellular matrix within human bone limits the applicability of conventional protocols for protein extraction. As a result, complete and accurate characterization of human bone proteome is yet to be performed, and as a result, several bone-related diseases such as craniosynostosis and osteosarcoma are still poorly understood. We sought to develop a reproducible method for extracting whole proteins of varying molecular weights from human cranial bone. Whole protein was extracted from human cranial suture by mechanically processing samples using a method that minimized heat introduction to proteins to limit degradation. Western blotting suggests the presence of whole protein while mass spectrometry was used to sequence peptides and identify isolated proteins. Molecular weights of extracted protein ranged from 9.4-629 kDa and contained proteins of both intra- and extra-cellular origin. High correlation scores among suture protein spectral counts suggest the reproducibility of the method. Ontology analytics revealed proteins of myriad functions including mediators of metabolic processes and cell organelles. These results suggest a reproducible method for isolation of whole protein representing a large range of molecular weights, origins and functions.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Dr. Russell R. Reid, MD, PhD

PROVIDER: MSV000080697 | MassIVE | Tue Mar 28 20:52:00 BST 2017

SECONDARY ACCESSION(S): PXD003215

REPOSITORIES: MassIVE

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A method for whole protein isolation from human cranial bone.

Lyon Sarah M SM Mayampurath Anoop A Rogers M Rose MR Wolfgeher Donald J DJ Fisher Sean M SM Volchenboum Samuel L SL He Tong-Chuan TC Reid Russell R RR

Analytical biochemistry 20160925

The presence of the dense hydroxyapatite matrix within human bone limits the applicability of conventional protocols for protein extraction. This has hindered the complete and accurate characterization of the human bone proteome thus far, leaving many bone-related disorders poorly understood. We sought to refine an existing method of protein extraction from mouse bone to extract whole proteins of varying molecular weights from human cranial bone. Whole protein was extracted from human cranial su ...[more]

PMID: 27677936

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