Synthesis and conformational properties of protein fragments based on the Id family of DNA-binding and cell-differentiation inhibitors.
Id proteins are dominant negative regulators of the helix-loop-helix (HLH) transcription factors and are important during development, especially by preventing cell differentiation while inducing cell proliferation. In contrast, they are poorly expressed in healthy adults but are found in several tumor types. The Id HLH motif is responsible for the inhibitory activity, whereas not much is known about the role of the N- and C-termini. In the presented work, synthetic peptides reproducing the HLH, the N-terminal region, and the C-terminal region of the Id proteins were characterized by CD. The four HLH sequences built highly stable helical conformations, whereas the N- and C-termini were unstructured, with the exception of an alanine-rich fragment preceding the Id4 HLH motif. Deletion of the loop connecting the two helices led to helix destabilization for all four Id HLH peptides. In addition, modifications of the amino acid composition within the hydrophobic face of the helices of the Id1 HLH peptide induced conformational changes, mostly associated with loss of helix content. Moreover, a fragment containing the helix-2 and the C-terminus of the Id1 protein did not show any helical character. Therefore, both the helix propensity and stability of the HLH domain were shown to be strongly dependent on favorable interhelical contacts. In contrast, it is suggested that the regions beyond this domain could rather play a destabilizing role, for example, by increasing the flexibility of the folded protein.
Kiewitz
Sebastian D
SD
Fakultät für Chemie und Pharmazie, Universität Regensburg, Universitätsstr. 31, 93053 Regensburg, Germany.
Cabrele
Chiara
C
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D002454:Cell Differentiation; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D017124:Conserved Sequence; D019281:Dimerization; D018507:Gene Expression Regulation, Developmental; D018257:Helix-Loop-Helix Motifs; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D051798:Inhibitor of Differentiation Protein 1 / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics; D051797:Inhibitor of Differentiation Protein 2 / Q000138:chemical synthesis / Q000737:chemistry; D051796:Inhibitor of Differentiation Proteins / Q000138:chemical synthesis / Q000737:chemistry; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D017384:Sequence Deletion; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014157:Transcription Factors / Q000235:genetics*
D051798:Inhibitor of Differentiation Protein 1; D051797:Inhibitor of Differentiation Protein 2; D051796:Inhibitor of Differentiation Proteins; D010455:Peptides; D014157:Transcription Factors