Two subunits of the 55 K porcine zona pellucida glycoprotein family are immunologically distinct
Abstract
The 55K glycoprotein family (ZP3) of the porcine zona pellucida is comprised of two subunits of 46 K and 45 K which can be resolved by endo-..beta..-galactosidase digestion of ZP3 followed by reversed phase HPLC on Vydac C4 resin. Gel electrophoresis revealed that the 46 K component (EBDG..cap alpha..) is approx. 95% pure and the 45 K component (EBGD..beta..) is 100% pure. In the present study, these two subunits were evaluated immunologically by RIA. Under similar reaction protocols (chloramine-T iodination procedure) comparable specific activities were obtained for EBGD..cap alpha.. (33.06 +/- 7.5 ..mu..ci/..mu..gm), EBGD..beta.. (30.45 +/- 1.6) and ZP3 (26.3 +/- 1.3). Antibody (Ab) titration studies revealed that EBGD..cap alpha.. and ..beta.. are potent immunogens and /sup 125/I-EBGD..cap alpha.. showed minimal cross reactivity to EBGD..beta..-Ab (8% bound at 1:500 dilution), whereas, /sup 125/I-EBGD..beta.. showed a greater degree of cross reactivity to EBGD..cap alpha..-Ab (23% bound at 1:500 dilution). Maximum binding for the two labeled antigens against homologous Abs (1:500) was > 60%. Dose response studies revealed that in the /sup 125/I-EBGD..cap alpha.. vs EBGD..cap alpha.. -Ab system, the 50% intercept was 3.25 +/- 0.32 ng for EBGD..cap alpha.. and 472.43 +/- 30.26 ng for EBGD..beta.. (p < 0.01), whereas, inmore »
- Authors:
- Publication Date:
- Research Org.:
- Wayne State Univ., Detroit, MI
- OSTI Identifier:
- 5463459
- Report Number(s):
- CONF-8604222-
Journal ID: CODEN: FEPRA; TRN: 86-028318
- Resource Type:
- Conference
- Journal Name:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
- Additional Journal Information:
- Journal Volume: 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; GALACTOSIDASE; ENZYME ACTIVITY; GLUCOPROTEINS; IMMUNE REACTIONS; ANTIBODIES; ANTIGENS; DOSE-RESPONSE RELATIONSHIPS; ELECTROPHORESIS; IODINE 125; LIQUID COLUMN CHROMATOGRAPHY; SWINE; TRACER TECHNIQUES; ANIMALS; BETA DECAY RADIOISOTOPES; CARBOHYDRATES; CHROMATOGRAPHY; DAYS LIVING RADIOISOTOPES; DOMESTIC ANIMALS; ELECTRON CAPTURE RADIOISOTOPES; ENZYMES; GLYCOSYL HYDROLASES; HYDROLASES; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; MAMMALS; NUCLEI; O-GLYCOSYL HYDROLASES; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; PROTEINS; RADIOISOTOPES; SACCHARIDES; SEPARATION PROCESSES; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques
Citation Formats
Subramanian, M G, Yurewicz, E C, and Sacco, A G. Two subunits of the 55 K porcine zona pellucida glycoprotein family are immunologically distinct. United States: N. p., 1986.
Web.
Subramanian, M G, Yurewicz, E C, & Sacco, A G. Two subunits of the 55 K porcine zona pellucida glycoprotein family are immunologically distinct. United States.
Subramanian, M G, Yurewicz, E C, and Sacco, A G. 1986.
"Two subunits of the 55 K porcine zona pellucida glycoprotein family are immunologically distinct". United States.
@article{osti_5463459,
title = {Two subunits of the 55 K porcine zona pellucida glycoprotein family are immunologically distinct},
author = {Subramanian, M G and Yurewicz, E C and Sacco, A G},
abstractNote = {The 55K glycoprotein family (ZP3) of the porcine zona pellucida is comprised of two subunits of 46 K and 45 K which can be resolved by endo-..beta..-galactosidase digestion of ZP3 followed by reversed phase HPLC on Vydac C4 resin. Gel electrophoresis revealed that the 46 K component (EBDG..cap alpha..) is approx. 95% pure and the 45 K component (EBGD..beta..) is 100% pure. In the present study, these two subunits were evaluated immunologically by RIA. Under similar reaction protocols (chloramine-T iodination procedure) comparable specific activities were obtained for EBGD..cap alpha.. (33.06 +/- 7.5 ..mu..ci/..mu..gm), EBGD..beta.. (30.45 +/- 1.6) and ZP3 (26.3 +/- 1.3). Antibody (Ab) titration studies revealed that EBGD..cap alpha.. and ..beta.. are potent immunogens and /sup 125/I-EBGD..cap alpha.. showed minimal cross reactivity to EBGD..beta..-Ab (8% bound at 1:500 dilution), whereas, /sup 125/I-EBGD..beta.. showed a greater degree of cross reactivity to EBGD..cap alpha..-Ab (23% bound at 1:500 dilution). Maximum binding for the two labeled antigens against homologous Abs (1:500) was > 60%. Dose response studies revealed that in the /sup 125/I-EBGD..cap alpha.. vs EBGD..cap alpha.. -Ab system, the 50% intercept was 3.25 +/- 0.32 ng for EBGD..cap alpha.. and 472.43 +/- 30.26 ng for EBGD..beta.. (p < 0.01), whereas, in the /sup 125/I-EBGD..beta.. vs EBGD..beta..-Ab system the 50% intercept was 3.51 +/- 0.58 for EBGD..beta.. and 166.77 +/- 49.20 for EBGD..cap alpha.. (p < 0.01). No significant differences were observed in the slopes of the dose response curves. It is concluded that the two subunits of ZP3 possess distinct immunologic characteristics as evaluated by RIA.},
doi = {},
url = {https://www.osti.gov/biblio/5463459},
journal = {Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)},
number = ,
volume = 45:3,
place = {United States},
year = {Sat Mar 01 00:00:00 EST 1986},
month = {Sat Mar 01 00:00:00 EST 1986}
}