Novel "omics" approach for study of low-abundance, low-molecular-weight components of a complex biological tissue: regional differences between chorionic and basal plates of the human placenta.
Tissue proteomics has relied heavily on two-dimensional gel electrophoresis, for protein separation and quantification, then single protein isolation, trypsin digestion, and mass spectrometric protein identification. Such methods are predominantly used for study of high-abundance, full-length proteins. Tissue peptidomics has recently been developed but is still used to study the most highly abundant species, often resulting in observation and identification of dozens of peptides only. Tissue lipidomics is likewise new, and reported studies are limited. We have developed an "omics" approach that enables over 7,000 low-molecular-weight, low-abundance species to be surveyed and have applied this to human placental tissue. Because the placenta is believed to be involved in complications of pregnancy, its proteomic evaluation is of substantial interest. In previous research on the placental proteome, abundant, high-molecular-weight proteins have been studied. Application of large-scale, global proteomics or peptidomics to the placenta have been limited, and would be challenging owing to the anatomic complexity and broad concentration range of proteins in this tissue. In our approach, involving protein depletion, capillary liquid chromatography, and tandem mass spectrometry, we attempted to identify molecular differences between two regions of the same placenta with only slightly different cellular composition. Our analysis revealed 16 species with statistically significant differences between the two regions. Tandem mass spectrometry enabled successful sequencing, or otherwise enabled chemical characterization, of twelve of these. The successful discovery and identification of regional differences between the expression of low-abundance, low-molecular weight biomolecules reveals the potential of our approach.
Department of Chemistry and Biochemistry, Brigham Young University, BNSN C-212, Provo, UT, 84602, USA.
Komal
K
Kedia
Department of Chemistry and Biochemistry, Brigham Young University, BNSN C-212, Provo, UT, 84602, USA.
Caitlin A
CA
Nichols
Department of Chemistry, Utah Valley University, 800 W. University Parkway, Orem, UT, 84058, USA.
Craig D
CD
Thulin
Department of Chemistry and Biochemistry, Brigham Young University, BNSN C-212, Provo, UT, 84602, USA. swgraves@chem.byu.edu.
Steven W
SW
Graves
D000595:Amino Acid Sequence; D002824:Chorionic Villi / Q000737:chemistry* / Q000378:metabolism; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D003656:Decidua / Q000737:chemistry* / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000302:isolation & purification*; D010743:Phospholipids / Q000302:isolation & purification*; D011247:Pregnancy; D020543:Proteome / Q000302:isolation & purification*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods; D053719:Tandem Mass Spectrometry
D010455:Peptides; D010743:Phospholipids; D020543:Proteome
Capillary liquid chromatography–MS; Lipids; Low-molecular-weight proteins; Peptidomics; Placenta; Tissue proteomics