Carboxymethylation of the human estrogen receptor ligand-binding domain-estradiol complex: HPLC/ESMS peptide mapping shows that cysteine 447 does not react with iodoacetic acid

Experiments were carried out to determine the degree of solvent and reagent accessibility of the cysteines in the ligand-binding domain of the human estrogen receptor (hER LBD). The cysteine residues were alkylated when human ER LBD was present in its ligand (estradiol)-bound conformation. Direct el...

Full description

Saved in:

Bibliographic Details
Published in:	Steroids Vol. 61; no. 6; pp. 367 - 373
Main Authors:	Hegy, Gilbert B., Shackleton, Cedric H.L., Carlquist, Mats, Bonn, Thomas, Engström, Owe, Sjöholm, Pelle, Witkowska, H.Ewa
Format:	Journal Article
Language:	English
Published:	New York, NY Elsevier Inc 01-06-1996 Elsevier Science
Subjects:	Alkylation Amino Acid Sequence Binding Sites Biological and medical sciences Cell receptors Cell structures and functions Chromatography, High Pressure Liquid Cysteine - metabolism Estradiol - metabolism estrogen receptor Fundamental and applied biological sciences. Psychology Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors HPLC/ESMS Humans Iodoacetates - chemistry Iodoacetic Acid ligand-binding domain MALDI mass spectrometry Mass Spectrometry - methods Methylation Molecular and cellular biology Molecular Sequence Data Peptide Fragments - chemistry Peptide Fragments - metabolism Peptide Mapping Protein Conformation Protein Denaturation Receptors, Estrogen - chemistry Receptors, Estrogen - metabolism mass spectrometry estrogen receptor HPLC/ESMS MALDI ligand-binding domain Human Cartography Cysteine Estrogen Binding site Sex steroid hormone Derivatization Hormonal receptor
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Experiments were carried out to determine the degree of solvent and reagent accessibility of the cysteines in the ligand-binding domain of the human estrogen receptor (hER LBD). The cysteine residues were alkylated when human ER LBD was present in its ligand (estradiol)-bound conformation. Direct electrospray ionization mass spectrometry (ESMS) as well as liquid chromatography coupled with ESMS, and matrix-assisted laser ionization desorption time-of-flight mass spectrometry were used to determine the location and the yield of the derivatized residues after proteolysis with trypsin. We observed that the cysteine 447 was protected against alkylation under these conditions, whereas cysteines 381, 417, and 530 were fully derivatized.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0039-128X 1878-5867
DOI:	10.1016/0039-128X(96)00042-6