title: [PREPARATION OF HUMAN TISSUE PROTEIN EXTRACTS ENRICHED WITH THE SPHINGOMYELIN SYNTHASE 1].
abstract: Sphingomyelin synthase 1 (SMS 1) catalyzes sphingomyelin biosynthesis in eukaryotic cells. We previously studied the structure of the human SGMS1 gene, which encodes the enzyme and its numerous transcripts. The tissue-specific expression of the transcripts was also described. Analysis of the SMS1 protein expression in human tissues using immunoblotting of tissue extracts prepared in the RIPA (Radio Immuno-Precipitation Assay) buffer revealed a weak signal in renal cortex, testis, lung, and no signal in placenta and lymphatic node. In this work, a new method of preparation of the tissue protein extracts enriched with SMS1 was suggested. The method based on the consecutive extraction with a buffer containing 0.05 and 1 mg/ml of the Quillaja saponaria saponin allowed SMS1 to be detected in all tissues tested. The SMS1 content in the saponin extract of kidney cortex is about 12-fold higher compared to the RIPA extraction procedure.
authors.forename: O Yu
authors.initials: OY
authors.lastname: Sudarkina
authors.forename: L V
authors.initials: LV
authors.lastname: Dergunova
mesh_terms: D001671:Biochemistry / Q000379:methods*; D002021:Buffers; D005591:Chemical Fractionation; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007672:Kidney Cortex / Q000737:chemistry; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry*; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000737:chemistry*; D031990:Quillaja / Q000737:chemistry; D012503:Saponins / Q000737:chemistry; D014020:Tissue Extracts / Q000737:chemistry*; D017855:Transferases (Other Substituted Phosphate Groups) / Q000032:analysis* / Q000737:chemistry*
chemical_list: D002021:Buffers; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D012503:Saponins; D014020:Tissue Extracts; C455440:SGMS1 protein, human; D017855:Transferases (Other Substituted Phosphate Groups)