Mass spectrometry identification of covalent attachment sites of two related estrogenic ligands on human estrogen receptor alpha

The Journal of Steroid Biochemistry and Molecular Biology
H MattrasJ L Borgna

Abstract

A purified preparation of human estrogen receptor alpha (hERalpha) ligand-binding domain (LBD) involving mainly the Ser(309)Ala(569) (approximately 30%) and Ser(309)Ala(571) (approximately 63%) ER portions was used to identify the covalent attachment sites of two closely related estrogenic ER affinity labels 17alpha-bromoacetamidopropylestradiol (17BAPE(2)) and 17alpha-bromoacetamidomethylestradiol (17BAME(2)). To identify and quantify the electrophile covalent attachment sites, [(14)C]17BAPE(2)- and [(14)C]17BAME(2)-alkylated hLBD preparations were trypsinized and submitted to HPLC. In each case, two radioactive fractions were obtained. Mass spectrometry analyses of the two fractions showed signals, which closely matched the molecular masses of alkylated Cys(530)Lys(531) and Cys(417)Arg(434) hLBD tryptic peptides. The covalent attachment of the two electrophiles on hLBD was assigned to the S atoms of Cys(530) and Cys(417). However, the balance between Cys(530) and Cys(417) labeling markedly differed according to the affinity label used, with the Cys(530)/Cys(417) ratio being 2.1 for 17BAPE(2), and 20 for 17BAME(2). We attempted to interpret the covalent attachment of electrophiles by molecular modeling using the crystallograph...Continue Reading

References

Jul 25, 1989·Nucleic Acids Research·J A LeesM G Parker
Oct 1, 1980·Molecular and Cellular Endocrinology·J L Borgna, H Rochefort
Jun 23, 1995·Journal of Medicinal Chemistry·D el GarroujJ L Borgna
Sep 1, 1994·The Journal of Steroid Biochemistry and Molecular Biology·M SalomonssonJ Häggblad
Oct 11, 1994·Proceedings of the National Academy of Sciences of the United States of America·V CavaillèsM G Parker
Jan 1, 1994·Annual Review of Biochemistry·M J Tsai, B W O'Malley
Feb 1, 1996·Nature Structural Biology·J M WurtzH Gronemeyer
Jun 11, 1996·Proceedings of the National Academy of Sciences of the United States of America·G G KuiperJ A Gustafsson
Jan 1, 1996·Analytical Chemistry·M Wilm, M Mann
Oct 24, 1997·Nature·A M BrzozowskiM Carlquist
May 30, 1998·Proceedings of the National Academy of Sciences of the United States of America·D M TanenbaumP B Sigler
Mar 16, 2001·Structure·A C PikeM Carlquist
Jul 5, 2001·Protein Expression and Purification·S EilerM Ruff
Sep 7, 2002·Molecular and Cellular Biology·Anna N MoraitisCatherine C Thompson
Jan 28, 2006·The Journal of Steroid Biochemistry and Molecular Biology·Sigrid AliauJean-Louis Borgna

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.