A combined top‐down and bottom‐up MS approach for the characterization of hemoglobin variants in Rhesus monkeys

Abstract

This article was originally published in Proteomics 2010, 10, 3657–3668, DOI

Familial secondary erythrocytosis due to increased oxygen affinity is caused by destabilization of the T state of hemoglobin Brigham (α2β2Pro100Leu)

“…LC‐ESI MS data were obtained using a single quadrupole MS (Agilent HP 1100 MSD series). Hundred microliters of the sample was injected through a C 3 column and similar gradient conditions were used as reported elsewhere 49. Deconvolution of the multiply charged species was performed using HP Chemstation software on the peaks integrated between 35 and 45 min by specifying a molecular mass range of 10 to 30 kDa, 5% abundance cutoff and three minimum peaks.…”

The top‐down, middle‐down, and bottom‐up mass spectrometry approaches for characterization of histone variants and their post‐translational modifications

“…Tools such as MS‐Deconv , YADA , Thrash , and Xtract (Thermo) can be used for generating a list of monoisotopic peaks, but it is not clear how well these algorithms perform with ETD or ECD, where the radical and nonradical fragment ions could hinder interpretation. In fact, just a simple determination of a monoisotopic peak that is readily accomplished in bottom‐up proteomics remains challenging when done with larger peptides . Additionally, problems can arise when trying to average MS2 spectra to increase the S/N ratio (as is commonly done in top‐down analyses), particularly when online separations are used.…”