A method for whole protein isolation from human cranial bone.
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 suture by mechanically processing samples using a method that limits protein degradation by minimizing heat introduction to proteins. The presence of whole protein was confirmed by western blotting. Mass spectrometry was used to sequence peptides and identify isolated proteins. The data have been deposited to the ProteomeXchange with identifier PXD003215. Extracted proteins were characterized as both intra- and extracellular and had molecular weights ranging from 9.4 to 629 kDa. High correlation scores among suture protein spectral counts support the reproducibility of the method. Ontology analytics revealed proteins of myriad functions including mediators of metabolic processes and cell organelles. These results demonstrate a reproducible method for isolation of whole protein from human cranial bone, representing a large range of molecular weights, origins and functions.
The Pritzker School of Medicine, United States.
Sarah M
SM
Lyon
Computation Institute, United States; Center for Research Informatics, United States.
Anoop
A
Mayampurath
The Molecular Oncology Laboratory, Department of Orthopedic Surgery, University of Chicago Medicine, United States.
M Rose
MR
Rogers
Proteomics Core Laboratory, The University of Chicago, Chicago, IL 60637, United States.
Donald J
DJ
Wolfgeher
The Pritzker School of Medicine, United States.
Sean M
SM
Fisher
Computation Institute, United States; Center for Research Informatics, United States.
Samuel L
SL
Volchenboum
The Molecular Oncology Laboratory, Department of Orthopedic Surgery, University of Chicago Medicine, United States.
Tong-Chuan
TC
He
The Laboratory of Craniofacial Development and Biology, Section of Plastic and Reconstructive Surgery, University of Chicago Medicine, United States. Electronic address: rreid@surgery.bsd.uchicago.edu.
Russell R
RR
Reid
D000818:Animals; D017886:Durapatite / Q000737:chemistry; D006801:Humans; D051379:Mice; D020543:Proteome / Q000302:isolation & purification* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012886:Skull / Q000737:chemistry* / Q000378:metabolism
D020543:Proteome; D017886:Durapatite
Cranial bone; Extraction; Human; Method; Protein
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