Effect of nitration on prolactin activities

Andersen, Thomas T.; Zamierowski, Marilyn M.; Ebner, Kurt E.

doi:10.1016/0003-9861(79)90076-6

Cited by 12 publications

(1 citation statement)

References 22 publications

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“…4 Depending on the disorders and proteins, the abundance of 3-nitrotyrosine was found to be increased by factors of 2-100, and 3-nitrotyrosine/ tyrosine ratios ranging from below 10 mmol mol 21 up to 50 mmol mol 21 have been measured. 5 Protein nitration occurs upon oxidative stress and inflammation 6,7 as well as during biological aging, 8,9 and several enzymes [10][11][12][13] and hormones 14,15 are known to be inactivated by nitration. In recent studies we have shown that proteins can also be efficiently nitrated by air pollutants such as nitrogen dioxide and ozone at environmentally relevant concentration levels.…”

Section: Introductionmentioning

confidence: 99%

Comparison of nitrotyrosine antibodies and development of immunoassays for the detection of nitrated proteins

Franze

Weller

Pöschl

2004

Analyst

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Three monoclonal antibodies (mAb) and three polyclonal antibodies (pAb) have been characterized and compared with respect to their cross-reactivities and affinities for 3-nitrotyrosine, eight aromatic compounds with similar chemical structures, a peptide containing a single nitrotyrosine residue, and fourteen nitrated protein standards (bovine serum albumin, BSA) containing different numbers of nitrotyrosine residues per protein molecule (0.2 to 16.8). In indirect competitive immunoassays, mAb Alexis 39B6 exhibited the highest affinity for free 3-nitrotyrosine (10(6) L mol(-1)), while the pAb Oxis 24312 from sheep exhibited the highest affinities for nitrated proteins (up to 10(8) L mol(-1)). The apparent affinities determined in the indirect competitive assays were inversely correlated with the limits of detection (LOD) determined in one-sided immunoassays. With the sheep pAb minimum LOD on the order of 10 pmol L(-1) were achieved for highly nitrated proteins, corresponding to effective LOD on the order of 100 pmol L(-1) for nitrotyrosine residues. In the one-sided assays, however, the LOD for nitrated proteins increased proportionally with increasing background concentrations of native proteins in the investigated samples. Sandwich immunoassays combining pAb and mAb for selective enrichment and detection of nitrated proteins allowed to eliminate this native protein matrix effect and to achieve LOD on the order of 300 pmol L(-1) for highly nitrated proteins independent of native protein background concentrations.

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Section: Introductionmentioning

confidence: 99%

Comparison of nitrotyrosine antibodies and development of immunoassays for the detection of nitrated proteins

Franze

Weller

Pöschl

2004

Analyst

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Effects of Prolactin on Target Cells

Frawley¹,

Porter²,

Kineman³

1990

Neuroendocrine Perspectives

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Solubilization of prolactin receptor by a Zwitterionic detergent

Church

Ebner

1982

Experientia

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A prolactin receptor was purified from a solu-bilized preparation of mouse microsomal membranes by affinity chromatography. The receptors were solubilized with the detergent 3-[(3-cholamidopropyl)dimethylammonio]-l-propanesulfo-nate) (Chaps) a zwitterionic derivative of cholic acid. The affinity gel was prepared by coupling ovine prolactin to CH-Sepharose 4B. After extensive elution of the nonspecifically adsorbed proteins, the receptors were eluted with 4 M urea/i M NaCI/0.5% Chaps followed by 5 M MgCl2/0.5% Chaps. The eluted receptor appeared on NaDodSO4/polyacrylamide gels as a single major band of Mr 37,000. The purified receptor retained its specificity for lactogenic hormones and binds "25I-labeled ovine prolactin with aKa of 2-6 x 109 M-1. Prolactin (PRL) plays a key role in development and differentiation of normal mammary glands (1) and in mammary tumor-igenesis (2). The initial step in PRL action is the interaction of this hormone with its receptor in the membrane. In the past, most studies attempting to understand the mechanism of PRL action have required that the receptor be examined solely by its ability to bind to lactogenic hormone. From these binding data, the mode of regulation of the receptor itself has been inferred. However, these observations have been based on the functionality of the receptor molecule. To effectively probe the nature of regulation of the PRL receptor itself, it is most desirable to have specific antibodies directed against it. To this end, we undertook the purification of a PRL receptor, using mouse liver as the target tissue and membrane source. This tissue contains both somatogenic and lactogenic hormone receptors (3). Previous attempts at purification of the lactogenic hormone (or PRL) receptor from target tissues have utilized Triton X-100 as the solubilization agent (4, 5). Because PRL aggregates in the presence of this detergent (4), human growth hormone (hGH) had to be used as the binding ligand. As a result, these studies have been hampered by the ability of hGH to bind to both so-matogenic and lactogenic hormone receptors (4, 6-8). Recently (9), we succeeded in solubilizing an active PRL receptor from mouse liver microsomal membranes by using the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-pro-panesulfonate (Chaps), a zwitterionic derivative of cholic acid (10). The solubilized material had the same binding affinity and specificity as those of the membrane receptor (9). Because PRL does not aggregate in the presence of Chaps and, thus, can effectively be utilized as a binding ligand (9), we attempted the purification ofthe PRL receptor, using an affinity column prepared by conjugating ovine PRL (oPRL) to Sepha-rose beads. The purified material, which consists of a major Mr 37,000 peptide, retains its specificity for lactogenic hormones and binds oPRL with the same high affinity as the membrane-bound receptor. MATERIALS AND METHODS Materials. Chemicals were from the following sources: oPRL (NIH-P-S-14) and bovine growth hormone (bGH; NIH-GH-B-18) were from t...

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Effect of nitration on prolactin activities

Cited by 12 publications

References 22 publications

Comparison of nitrotyrosine antibodies and development of immunoassays for the detection of nitrated proteins

Comparison of nitrotyrosine antibodies and development of immunoassays for the detection of nitrated proteins

Effects of Prolactin on Target Cells

Solubilization of prolactin receptor by a Zwitterionic detergent

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