Membrane proteins of human fetal primitive nucleated red blood cells

• Published in: Journal of proteomics 2012-10, Vol.75 (18), p.5762-5773
• Main Authors: Ponnusamy, Sukumar, Zhang, Huoming, Kadam, Priya, Lin, Qingsong, Lim, Teck Kwang, Sandhu, Jaspal Singh, Kothandaraman, Narasimhan, Mahyuddin, Aniza Puteri, Biswas, Arijit, Venkat, Annapoorna, Hew, Choy-Leong, Joshi, Shashikant B., Chung, Maxey Ching Ming, Choolani, Mahesh
• Format: Article
• Language: English
• Subjects: 2D-LC-MALDITOF/TOF-MS; adults; bioinformatics; Biological and medical sciences; Diverse techniques; Erythrocytes; fetus; Fundamental and applied biological sciences. Psychology; Human fetal nucleated red blood cells; humans; immunocytochemistry; liquid chromatography; mass spectrometry; Membrane protein profiling; Molecular and cellular biology; Noninvasive prenatal diagnosis; plasma membrane; proteome; proteomics; RBC; reverse transcriptase polymerase chain reaction; surface antigens; transferrin; transmembrane proteins
• ISSN: 1874-3919
• DOI: 10.1016/j.jprot.2012.07.020

In humans, primitive fetal nucleated red blood cells (FNRBCs) are thought to be as vital for embryonic life as their counterpart, adult red blood cells (adult RBCs) are in later-gestation fetuses and adults. Unlike adult RBCs, the identity and functions of FNRBC proteins are poorly understood owing to a scarcity of FNRBCs for proteomic investigations. The study aimed to investigate membrane proteins of this unique cell type. We present here, the first report on the membrane proteome of human primitive FNRBCs investigated by two-dimensional liquid chromatography coupled with mass-spectrometry (2D-LCMS/MS) and bioinformatics analysis. A total of 273 proteins were identified, of which 133 (48.7%) were membrane proteins. We compared our data with membrane proteins of adult RBCs to identify common, and unique, surface membrane proteins. Twelve plasma membrane proteins with transmembrane domains and eight proteins with transmembrane domains but without known sub-cellular location were identified as unique-to-FNRBCs. Except for the transferrin receptor, all other 19 unique-to-FNRBC membrane proteins have never been described in RBCs. Reverse-transcriptase PCR (RT-PCR) and immunocytochemistry validated the 2D-LCMS/MS data. Our findings provide potential surface antigens for separation of primitive FNRBCs from maternal blood for noninvasive prenatal diagnosis, and to understand the biology of these rare cells. [Display omitted] ► First report on membrane proteome of human fetal red blood cells (FNRBCs) ► Unique-to-FNRBC plasma membrane proteins compared to adult RBCs are identified. ► Potential surface-antigens for enrichment of FNRBCs from maternal blood are identified. ► RT-PCR and Immunocytochemistry validated 2DLCMS/MS identifications. ► 171 surface proteins of adult RBCs are not present in FNRBCs.