A Comparative Protein Profile of Mammalian Erythrocyte Membranes Identified by Mass Spectrometry

Abstract

A comparative analysis of erythrocyte membrane proteins of economically important animals, goat (Capra aegagrus hircus), buffalo (Bubalus bubalis), pig (Sus scrofa), cow (Bos tauras), and human (Homo sapiens) was performed. Solubilized erythrocyte membrane proteins were separated by sodium dodecyl sulfate-polyacryamide gel electrophoresis (SDS-PAGE), visualized by staining the gels with Commassie Brilliant Blue (CBB), and identified by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Emerging results show that all major erythrocyte membrane proteins present in human are also seen in all the animals except for band 4.5 which could not be identified. Band 3 is seen as more intense and compact, band 4.1 appears as a doublet in all the animal erythrocyte membranes, band 4.2 exhibits a slightly higher molecular weight (Mr) in buffalo, and cow and band 4.9 has a higher Mr in all the animals relative to the human protein. In addition, there are two new bands in the goat membrane, band G1, identified as HSP 90α, and band G2 identified as HSP 70. A new band C2 identified as HSP 70 is also seen in cow membranes. Peroxiredoxin II is of lower intensity and/or higher Mr in the animals. The difference in size of the proteins possibly indicates the variations in the composition of the amino acids. The difference in intensity of the proteins among these mammalians highlights the presence of less or more number of copies of that protein per cell. This data complement the earlier observations of differences in the sialoglycoprotein profile and effect of proteases and neuraminidase on agglutination among the mammalian erythrocytes. This study provides a platform to understand the molecular architecture of the individual erythrocytes, and in turn the dependent disorders, their phylogenetic relationship and also generates a database of erythrocyte membrane proteins of mammals. The animals selected for this study are of economic importance as they provide milk for the dairy industry and raw material for leather industry and are routinely sacrificed to obtain non vegetarian food worldwide.

Modifications of proteins of membrane-cytoskeleton complex and production of reactive oxygen species in erythrocytes cryopreserved with polyethylene glycol

“…~60 kDa and band 8 (Fig. 1A, lane 2), which have recently been identified as catalase and peroxiredoxin [10]. Dissociation of these proteins from MCC may indicate the activation of ROS production in destroyed cells.…”

“…The protein band 8 is peroxiredoxin 2 [10], which takes part in the cell protection from oxidative stress [13]. It contains a functional, readily available −SH-group of cysteine that explains its disappearance from the protein profile of control erythrocytes due to its transition to the HMWPC under the diamide effect.…”

The effect of cryoprotective agents on proteins of the erythrocyte membrane-cytoskeleton complex

“…В использованных экспериментальных условиях (концентрация диамида и условия инкубации) лишь незначительная часть спектрина, а также белков полос 3 и 5 включалась в образование ВПК в контрольных клетках [2]. Вместе с тем, белок с молекулярной массой 60 кДа и белок полосы 8, представляющие собой каталазу и пероксиредоксин 2 [10], практически полностью исчезали из электрофоретического профиля, что обусловлено легкой доступностью их функциональных -SH-групп для диамида.…”

Comparative Analysis of Erythrocyte Proteomes of Water Buffalo, Dairy Cattle, and Beef Cattle by Shotgun LC-MS/MS

“…Proteomics, an emerging technology, refers to the protein-expression profiles of a gene, a cell, or a tissue in a particular period (5, 20–22). Unlike the immutable genomics, proteomics is a post-genomic method and is preferentially sensitive to dynamic changes in the parasite and the host; that is, it changes along with the environment.…”

“…For instance, proteomics is used in the analysis of female Rhipicephalus Microplus -stage-specific protein expression of B. bovis and also in finding biomarkers for the diagnosis of Babesia canis (21, 24). In addition, a protein profile of mammalian erythrocyte membranes has been identified by matrix-assisted laser desorption/ionization time-of-flight/mass spectrometer (MALDI-TOF/MS) (22). Proteomics has also been applied to develop vaccines against tick-borne diseases (5).…”