Asparagine deamidation reduces DNA‐binding affinity of theDrosophila melanogaster Scr homeodomain. Issue 21 (3rd October 2015)

• Record Type: Journal Article
• Authors: O'Connell, Nichole E.; Lelli, Katherine; Mann, Richard S.; Palmer, Arthur G.
• Is Part Of: FEBS letters. Volume 589:Issue 21(2015)
• Page Start: 3237
• Page End: 3241
• Publication Date: 2015-10-03
• Subjects: Asparagine deamidation -- Dissociation constant -- DNA binding -- Hox transcription factor -- Sex Combs Reduced -- NMR spectroscopy; Biochemistry -- Periodicals; Biophysics -- Periodicals; Molecular biology -- Periodicals; Biochimie -- Périodiques; Biochemistry; Biophysics; Molecular biology; Periodicals; 572.05
• DOI: 10.1016/j.febslet.2015.09.020
• Languages: English
• ISSNs: 0014-5793

Abstract

Abstract : Spontaneous deamidation of asparagine is a non‐enzymatic post‐translational modification of proteins. Residue Asn 321 is the main site of deamidation of the Drosophila melanogaster Hox transcription factor Sex Combs Reduced (Scr). Formation of iso‐aspartate, the major deamidation product, is detected by HNCACB triple‐resonance NMR spectroscopy. The rate of deamidation is quantified by fitting the decay of Asn NH2 side‐chain signals in a time‐series of 15 N‐ 1 H HSQC NMR spectra. The deamidated form of Scr binds to specific DNA target sequences with reduced affinity as determined by an electrophoretic mobility shift assay. Abstract : Drosophila melanogaster Hox transcription factor Sex Combs Reduced (Scr) is spontaneously deamidated at residue Asn 321. NMR spectroscopy is used to detect and quantify deamidation. Deamidation reduces the affinity of Scr for target DNA.