title: Conformational adjustments of SNase R and its N-terminal fragments probed by monoclonal antibodies.
abstract: Two monoclonal antibodies specific for staphylococcal nuclease R (SNase R) (McAb2C9 and McAb1B8) were prepared and used to probe protein folding during peptide elongation, by measuring antibody binding to seven N-terminal fragments (SNR141, SNR135, SNR121, SNR110, SNR102, SNR79 and SNR52) of SNase R. Comparative studies of the conformations of the N-terminal fragments have shown that all seven fragments of SNase R have a certain amount of residual structure, indicating that folding may occur during elongation of the nascent peptide chain. We show that the binding abilities of the intact enzyme and its seven fragments to the monoclonal antibodies are not simply proportional to the length of the peptide chain, suggesting that there may be continuous conformational adjustment in the nascent peptide chain as new C-terminal amino acids are added. A folding intermediate close in structure to the native state but with structural features in common with SNR121 is highly populated in 0.6 M GuHCl, and is also formed transiently during folding.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Academia Sinica, Beijing, PR China.
authors.forename: Z J
authors.initials: ZJ
authors.lastname: Qin
authors.forename: Y Q
authors.initials: YQ
authors.lastname: Tang
authors.forename: G Z
authors.initials: GZ
authors.lastname: Jing
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Li
authors.forename: G C
authors.initials: GC
authors.lastname: Huang
authors.forename: S
authors.initials: S
authors.lastname: Perrett
authors.forename: Z Q
authors.initials: ZQ
authors.lastname: Guo
authors.forename: J M
authors.initials: JM
authors.lastname: Zhou
mesh_terms: D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D007074:Immunoglobulin G; D008836:Micrococcal Nuclease / Q000737:chemistry*; D010441:Peptide Chain Elongation, Translational; D010446:Peptide Fragments / Q000737:chemistry* / Q000276:immunology; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D010446:Peptide Fragments; C104574:staphylococcal nuclease R; D008836:Micrococcal Nuclease