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Biochemical Studies of Human Placental Microvillous Plasma Membrane Proteins
| Content Provider | Scilit |
|---|---|
| Author | Webb, P. D. Evans, P. W. Molloy, C. M. Johnson, P. M. |
| Copyright Year | 1985 |
| Description | Journal: American Journal of Reproductive Immunology Isolated human syncytiotrophoblast microvillous plasma membranes (StMPM) have been examined by electron microscopy, SDS-polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional PAGE (2D-PAGE), and immunoblots. Electron microscopy of StMPM pellets revealed populations of membrane-bounded vesicles that disrupted after treatment with the chaotrope 3M KCl for 16 hr; with increasing molarity of another chaotrope (NH4SCN), the vesicles became smaller and more homogeneous. NH4SCN treatment resulted in significant reduction on SDS and 2D-PAGE analysis of only one protein at 80kd, shown by immunoblotting to be transferrin; 3M KCl had little effect and appeared to be a poor chaotrope. Chromogenic silver staining of SDS-PAGE gels demonstrated over 50 StMPM-associated discrete protein components. Immunoblotting revealed transferrin (80kd), albumin (65kd), IgG heavy chain (56kd), and Gc protein (56kd). Alpha-2-macroglobulin (alpha 2M) was identified at 180kd and 95kd; the smaller component may be a proteolytic derivative indicating alpha 2M binding to a trophoblast surface protease. Numerous discrete protein dots, and groups of dots characteristic of charge heterogeneity of individual proteins, were observed on high resolution 2D-PAGE. The most intensely stained proteins were transferrin (80kd), albumin (65kd), placental-type alkaline phosphatase (66kd), and actin (46kd). This 2D-PAGE technique is a superior method for analyzing the trophoblast membrane proteins, and the system described will enable systematic mapping of these components. |
| Related Links | http://onlinelibrary.wiley.com/doi/10.1111/j.1600-0897.1985.tb00321.x/pdf |
| Ending Page | 119 |
| Page Count | 7 |
| Starting Page | 113 |
| e-ISSN | 16000897 |
| DOI | 10.1111/j.1600-0897.1985.tb00321.x |
| Journal | American Journal of Reproductive Immunology |
| Issue Number | 4 |
| Volume Number | 8 |
| Language | English |
| Publisher | Wiley-Blackwell |
| Publisher Date | 1985-08-01 |
| Access Restriction | Open |
| Subject Keyword | Journal: American Journal of Reproductive Immunology Dentistry and Oral Surgery Syncytiotrophoblast Membrane Two‐dimensional Polyacrylamide Gel Electrophoresis |
| Content Type | Text |
| Resource Type | Article |
