通过位点特异性金属催化氧化对人催乳素金属结合位点的表征

Characterization of the metal-binding site of human prolactin by site-specific metal-catalyzed oxidation.

作者信息

Sadineni Vikram, Galeva Nadezhda A, Schöneich Christian

机构信息

Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS 66047, USA.

出版信息

Anal Biochem. 2006 Nov 15;358(2):208-15. doi: 10.1016/j.ab.2006.08.026. Epub 2006 Sep 12.

DOI:10.1016/j.ab.2006.08.026

PMID:17010299

Abstract

Site-specific metal-catalyzed oxidation (MCO) was applied to characterize the metal-binding site (MBS) of recombinant human prolactin (hPRL), which belongs to the hematopoietic cytokine family. Copper and ascorbate of various concentrations were used to initiate the oxidation of hPRL, and the oxidation-sensitive motifs were characterized and quantitated by mass spectrometry. Based on the results obtained with 10 microM Cu(2+) and 0.3-2.0mM ascorbate, we propose that the MBS in hPRL is composed of His27, His30, and His173. This result shows the similarity of hPRL to human growth hormone (hGH), a member of the same family as hPRL, where the MBS is composed of His18, His21, and Glu174.

摘要

位点特异性金属催化氧化（MCO）被用于表征重组人催乳素（hPRL）的金属结合位点（MBS），hPRL属于造血细胞因子家族。使用不同浓度的铜和抗坏血酸引发hPRL的氧化，并通过质谱对氧化敏感基序进行表征和定量。基于用10 microM Cu(2+)和0.3 - 2.0 mM抗坏血酸获得的结果，我们提出hPRL中的MBS由His27、His30和His173组成。该结果显示了hPRL与人生长激素（hGH）的相似性，hGH与hPRL属于同一家族，其MBS由His18、His21和Glu174组成。