Biochemical characterization and crystalization of human Zn-α2-glycoprotein, a soluble class I major histocompatibility complex homolog
Abstract
Zn-α2-glycoprotein (ZAG) is a 41-kDa soluble protein that is present in most bodily fluids. In addition, ZAG accumulates in fluids from breast cysts and in 40% of breast carcinomas, which suggests that ZAG plays a role in the development of breast diseases. However, the function of ZAG under physiological and cancerous conditions remains unknown. Because ZAG shares 30–40% sequence identity with the heavy chains of class I major histocompatibility complex (MHC) proteins, we compared the biochemical properties of ZAG with those of classical class I MHC molecules. We purified human ZAG from breast cyst fluid and serum and produced a panel of anti-ZAG monoclonal antibodies. Binding assays and acid elution experiments revealed that, in contrast to class I MHC proteins, ZAG does not bind peptides or the class I light chain, β2-microglobulin (β_(2)m). Nevertheless, CD studies indicated that ZAG is thermally stable in the absence of bound peptide or associated β2m, as opposed to class I MHC molecules, which require the presence of both β2m and peptides for stability. These data indicate that the function of ZAG has diverged from the peptide presentation and T-cell interaction functions of class I molecules. To gain insight into the function of ZAG and to compare the three-dimensional structures of ZAG and class I MHC molecules, we produced ZAG crystals that diffract beyond 2.7 Å and have initiated an x-ray structure determination.
Additional Information
© 1997 The National Academy of Sciences. Received December 16, 1996; accepted February 21, 1997. We thank Dr. F. Vizoso for breast cyst fluid, Dr. S. Mayo and his laboratory for use of the CD spectrometer, Dr. M. Harrington for use of the PhastSystem, S. Ou and the Caltech monoclonal antibody facility, Dr. Gary Hathaway and the Caltech PPMAL facility, and Dr. A. Chirino for data collection and processing at Cornell High Energy Synchrotron Source. L.M.S. was supported by the Fulbright Visiting Scholar Program.Attached Files
Published - PNAS-1997-Sánchez-4626-30.pdf
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Additional details
- PMCID
- PMC20774
- Eprint ID
- 52316
- Resolver ID
- CaltechAUTHORS:20141203-095910857
- Fulbright Foundation
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2014-12-03Created from EPrint's datestamp field
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2021-11-10Created from EPrint's last_modified field