title: Conformational stability of six truncated cHMG1a proteins studied in their mixture by H/D exchange and electrospray ionization mass spectrometry.
abstract: The high mobility group (HMG) proteins are abundant non-histone components of eukaryotic chromatin. The presence of C-terminal acidic tails is a common feature of the majority of HMG proteins. Although the biological significance of the acidic domains is not clear, they are conferring conformational and metabolic stability to the proteins in vitro. Moreover, the length and net charge of the acidic tails affect the strength of HMG protein interaction with DNA. Synthesis of an insect HMG protein by standard recombinant technology in bacteria leads to a mixture of the intact protein (cHMG1a-(1-113) (I)) and a series of its degradation products truncated at the C tail: cHMG1a-(1-111) (II); cHMG1a-(1-110) (III); cHuMGla-(1-109) (IV); cHMG1a-(1-108) (V); cHMG1a-(1-107) (VI); cHMG1a-(1-106) (VII). The proteins differ from each other only by the number of amino-acid residues at the C-terminal tail. We used H/D exchange mass spectrometry to characterize the stability of the proteins directly in their mixture. The results show that the proteins I-V and VII have very similar conformations. The protein VI is less compact and exchanges its protons faster than the others. It may be concluded that the C-terminal tail influences the conformation of the cHMG1a protein and that individual residues in this part of the protein play a key role in its compactness.
authors: Faculty of Chemistry, University of Wrocław, Poland. inga@wchuwr.chem.uni.wroc.pl
authors: I
authors: I
authors: Petry
authors: J R
authors: JR
authors: Wigniewski
authors: Z
authors: Z
authors: Szewczuk
mesh_terms: D003001:Cloning, Molecular; D024243:HMGB1 Protein / Q000737:chemistry* / Q000235:genetics; D013058:Mass Spectrometry; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D021241:Spectrometry, Mass, Electrospray Ionization; D013816:Thermodynamics; D013997:Time Factors
chemical_list: D024243:HMGB1 Protein