The citation is:
Philipp F Lange, Pitter F Huesgen, Karen Nguyen, and Christopher M Overall. " Annotating N termini for the Human Proteome Project: N termini and N#-acetylation status differentiate stable cleaved protein species from degradation remnants in the human erythrocyte proteome”, J. Proteome Research., Just Accepted Manuscript • DOI: 10.1021/pr401191w • Publication Date (Web): 21 Feb 2014.
In brief, the article describes a goal of the Chromosome-centric Human Proteome Project to identify all human protein species. With 3,844 proteins annotated as “missing” this is challenging. Enucleated and largely void of internal membranes and organelles, erythrocytes are simple yet proteomically challenging cells due to the high hemoglobin content (about 97% by mass) and wide dynamic range of protein concentrations that impedes protein identification. Using a N-terminomics procedure called TAILS, the authors identified 1369 human erythrocyte natural and neo-N-termini and 1234 proteins. From the HemoVoid™ treated, hemoglobin-depleted soluble fraction, 778 proteins were identified, 171of which were not represented in either the soluble non-depleted fraction or the membrane fraction. This study also establishes a general workflow suitable for the in-depth determination of the position and nature of human protein N termini in different tissues and disease states. The identification of 281 novel erythrocyte proteins and six missing proteins identified for the first time in the human proteome confirmed its utility.
“While the authors acknowledged other low abundance enrichment methods, our HemoVoid™ product was chosen for protein level enrichment. I am pleased to see it proved exceedingly useful in this exciting new area of proteomic identification.”
HemoVoid™ , is a proprietary porous silica-based protein enrichment matrix. It is designed to substantially deplete hemoglobin from erythrocyte lysates while concurrently enriching the low abundance proteome without bias towards pI or molecular weight. The HemoVoid™ protocol uses mild buffers so native enzyme activity is retained in elution fractions.
Key features are:
Hemoglobin voids in flow-through >98%
<30 minute bind/wash/elute protocol
Ideal for erythrocyte proteomics
Depletion from heavily hemolyzed serum, blood and dried blood spot/blood card
Applicable to hemoglobin variant analysis
Low abundance protein and enzyme enrichment
Mild conditions maintain biological or enzymatic activity
Native structure uncompromised with simple transfer to post-treatment interrogations
Species and tissue agnostic
Compatible with LC-MS, activity-probe profiling and virtually all proteomic analyses
On-bead trypsin digestion protocols for simple, consistent LC-MS analysis
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About Biotech Support Group LLC
Biotech Support Group LLC is a leading developer of proteomic, metabolomic and genomic sample preparation and enrichment products. It’s principal products include: AlbuVoid™ & AlbuSorb™ for albumin depletion, Cleanascite™
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