Circular Dichroic Analysis of Denatured Proteins: Inclusion of Denatured Proteins in the Reference Set

Venyaminov, Sergei Yu.; Baikalov, Igor; Shen, Zhi Min; Wu, C.S.C.; Yang, Jen Tsi

doi:10.1006/abio.1993.1450

Cited by 121 publications

(84 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Interestingly, in the CD spectra of the PAS sequences the absence of a weak positive maximum around 225 nm, which was often seen in other studies for polypeptides in random‐coil conformation having more average amino acid composition,44 is reminiscent of the “unordered poly‐proline structure” described by Tiffany and Krimm40 that is characterized by a strong minimum around 200 nm only (Supporting Information Table S3). Taking into consideration that repetitive amino acid sequences are unlikely to adopt an ideal random conformation without any (at least locally) regular structure,45 it would be plausible that PAS polypeptides show a kind of random‐coil structure influenced by the PPII conformation 46.…”

Section: Resultssupporting

confidence: 59%

The polypeptide biophysics of proline/alanine‐rich sequences (PAS): Recombinant biopolymers with PEG‐like properties

Breibeck

Skerra

2017

Biopolymers

View full text Add to dashboard Cite

PAS polypeptides comprise long repetitive sequences of the small L‐amino acids proline, alanine and/or serine that were developed to expand the hydrodynamic volume of conjugated pharmaceuticals and prolong their plasma half‐life by retarding kidney filtration. Here, we have characterized the polymer properties both of the free polypeptides and in fusion with the biopharmaceutical IL‐1Ra. Data from size exclusion chromatography, dynamic light scattering, circular dichroism spectroscopy and quantification of hydrodynamic and polar properties demonstrate that the biosynthetic PAS polypeptides exhibit random coil behavior in aqueous solution astonishingly similar to the chemical polymer poly‐ethylene glycol (PEG). The solvent‐exposed PAS peptide groups, in the absence of secondary structure, account for strong hydrophilicity, with negligible contribution by the Ser side chains. Notably, PAS polypeptides exceed PEG of comparable molecular mass in hydrophilicity and hydrodynamic volume while exhibiting lower viscosity. Their uniform monodisperse composition as genetically encoded polymers and their biological nature, offering biodegradability, render PAS polypeptides a promising PEG mimetic for biopharmaceutical applications.

show abstract

Section: Resultssupporting

confidence: 59%

The polypeptide biophysics of proline/alanine‐rich sequences (PAS): Recombinant biopolymers with PEG‐like properties

Breibeck

Skerra

2017

Biopolymers

View full text Add to dashboard Cite

show abstract

“…Fitted parameters were the melting temperature T M , at which 50% of proteins are in the folded and unfolded state, van't Hoff's enthalpy ΔH vH at the transition midpoint and the slope and Θ‐intercept of the native baseline assumed as a linear function of the temperature. As all protein complexes aggregated at various degrees of unfolding, the ellipticity of the unfolded state was set as a constant of −4400 degrees cm 2 dmol −1 ; this value resulted from fitting melting curves of LMP2A LTE and positive control peptide VTEHDTLLY‐containing complexes, which showed the least aggregation tendency, and is in good agreement with values reported for other thermally denatured proteins 34. For all peptides, the coefficient of determination for fitted curves versus measurements was r 2 > 0·99.…”

Section: Methodssupporting

confidence: 54%

The impact of HLA class I and EBV latency-II antigen-specific CD8+ T cells on the pathogenesis of EBV+ Hodgkin lymphoma

Jones

Wockner

Brennan

et al. 2015

Clinical and Experimental Immunology

View full text Add to dashboard Cite

SummaryIn 40% of cases of classical Hodgkin lymphoma (cHL), Epstein–Barr virus (EBV) latency‐II antigens [EBV nuclear antigen 1 (EBNA1)/latent membrane protein (LMP)1/LMP2A] are present (EBV+cHL) in the malignant cells and antigen presentation is intact. Previous studies have shown consistently that HLA‐A*02 is protective in EBV+cHL, yet its role in disease pathogenesis is unknown. To explore the basis for this observation, gene expression was assessed in 33 cHL nodes. Interestingly, CD8 and LMP2A expression were correlated strongly and, for a given LMP2A level, CD8 was elevated markedly in HLA‐A*02– versus HLA‐A*02+ EBV+cHL patients, suggesting that LMP2A‐specific CD8+ T cell anti‐tumoral immunity may be relatively ineffective in HLA‐A*02– EBV+cHL. To ascertain the impact of HLA class I on EBV latency antigen‐specific immunodominance, we used a stepwise functional T cell approach. In newly diagnosed EBV+cHL, the magnitude of ex‐vivo LMP1/2A‐specific CD8+ T cell responses was elevated in HLA‐A*02+ patients. Furthermore, in a controlled in‐vitro assay, LMP2A‐specific CD8+ T cells from healthy HLA‐A*02 heterozygotes expanded to a greater extent with HLA‐A*02‐restricted compared to non‐HLA‐A*02‐restricted cell lines. In an extensive analysis of HLA class I‐restricted immunity, immunodominant EBNA3A/3B/3C‐specific CD8+ T cell responses were stimulated by numerous HLA class I molecules, whereas the subdominant LMP1/2A‐specific responses were confined largely to HLA‐A*02. Our results demonstrate that HLA‐A*02 mediates a modest, but none the less stronger, EBV‐specific CD8+ T cell response than non‐HLA‐A*02 alleles, an effect confined to EBV latency‐II antigens. Thus, the protective effect of HLA‐A*02 against EBV+cHL is not a surrogate association, but reflects the impact of HLA class I on EBV latency‐II antigen‐specific CD8+ T cell hierarchies.

show abstract

“…2) correlate well with the information provided by pH-potentiometric and UV data. The spectrum measured at pH = 4.65 is very similar to that of the free ligand at the same pH (see 3) [53,54]. Spectra measured in the absence and presence of zinc(II) at pH = 3 are almost identical indicating no complex formation at this pH (spectra not shown).…”

Section: Spectroscopic Studies On the Zinc(ii) -Peptide Complexesmentioning

confidence: 63%

Competition of zinc(II) with cadmium(II) or mercury(II) in binding to a 12-mer peptide

Jancsó

Gyurcsik

Mesterházy

et al. 2013

Journal of Inorganic Biochemistry

View full text Add to dashboard Cite

show abstract

Circular Dichroic Analysis of Denatured Proteins: Inclusion of Denatured Proteins in the Reference Set

Cited by 121 publications

References 0 publications

The polypeptide biophysics of proline/alanine‐rich sequences (PAS): Recombinant biopolymers with PEG‐like properties

The polypeptide biophysics of proline/alanine‐rich sequences (PAS): Recombinant biopolymers with PEG‐like properties

The impact of HLA class I and EBV latency-II antigen-specific CD8+ T cells on the pathogenesis of EBV+ Hodgkin lymphoma

Competition of zinc(II) with cadmium(II) or mercury(II) in binding to a 12-mer peptide

Contact Info

Product

Resources

About