Measuring and Increasing Protein Solubility

Treviño, Saul; Scholtz, J. Martin; Pace, C. Nick

doi:10.1002/jps.21327

Cited by 134 publications

(135 citation statements)

References 94 publications

Supporting

Mentioning

129

Contrasting

Unclassified

Order By: Relevance

“…This hypothesis was validated by the analysis of the solubility-changing substitutions in proteins, which revealed that serine, together with glutamic acid and aspartic acid, contributes more favorably to protein solubility than other hydrophilic residues (asparagine, glutamine, threonine, lysine, and arginine). 95 …”

Section: Biological Significance Of Free Serinementioning

confidence: 99%

The intrinsic disorder alphabet. III. Dual personality of serine

Uversky

2015

Intrinsically Disordered Proteins

View full text Add to dashboard Cite

Proteins are natural polypeptides consisting of 20 major amino acid residues, content and order of which in a given amino acid sequence defines the ability of a related protein to fold into unique functional state or to stay intrinsically disordered. Amino acid sequences code for both foldable (ordered) proteins/domains and for intrinsically disordered proteins (IDPs) and IDP regions (IDPRs), but these sequence codes are dramatically different. This difference starts with a very general property of the corresponding amino acid sequences, namely, their compositions. IDPs/IDPRs are enriched in specific disorder-promoting residues, whereas amino acid sequences of ordered proteins/domains typically contain more order-promoting residues. Therefore, the relative abundances of various amino acids in ordered and disordered proteins can be used to scale amino acids according to their disorder promoting potentials. This review continues a series of publications on the roles of different amino acids in defining the phenomenon of protein intrinsic disorder and represents serine, which is the third most disorder-promoting residue. Similar to previous publications, this review represents some physico-chemical properties of serine and the roles of this residue in structures and functions of ordered proteins, describes major posttranslational modifications tailored to serine, and finally gives an overview of roles of serine in structure and functions of intrinsically disordered proteins.

show abstract

Section: Biological Significance Of Free Serinementioning

confidence: 99%

The intrinsic disorder alphabet. III. Dual personality of serine

Uversky

2015

Intrinsically Disordered Proteins

View full text Add to dashboard Cite

show abstract

“…These typical studies include antibody primary structure by chemical sequencing (Edman degradation) as well as peptide mapping by means of ESI-MS and MALDI-MS [26][27][28]; higher order structure by RP-HPLC-ESI-MS, Ellman's assay CD, FTIR, hydrogen deuterium exchange (HDX)-MS and X-ray [28,29]; post-translational modifications (PTM) and charge variants by MSion-exchange chromatography (IEC), hydrophobic interaction chromatography (HIC), capillary electrophoresis (CE) and isoelectric focusing (IEF) [30]; glycan profile and variants and size heterogeneity by HPLC-fluorescence, size-exclusion chromatography (SEC), native gel, capillary electrophoresis, RP-HPLC-MS and native intact MS [31], as well as solubility measurement by ultrafiltration or PEG-induced precipitation methods [32,33]. Moreover, the immunogenicity should be evaluated for both antibody and ADCs [34,35], but unnecessary for SMs.…”

Section: General Differences Between Sms and Lms (Mab And Adc) In Admmentioning

confidence: 99%

An Overall Comparison of Small Molecules and Large Biologics in ADME Testing

Wan

2016

ADMET DMPK

View full text Add to dashboard Cite

Biologics mainly monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) as new therapeutics are becoming increasingly important biotherapeutics. This review is intended to provide an overall comparison between small molecules (SMs) and biologics or large molecules (LMs) concerning drug metabolism and pharmacokinetic (DMPK) or associated with absorption, distribution, metabolism and elimination (ADME) testing from pharmaceutical industry drug discovery and development points of view, which will help design and conduct relevant ADME testing for biologics such as mAbs and ADCs. Recent advancements in the ADME for testing biologics and related bioanalytical methods are discussed with an emphasis on ADC drug development as an example to understand its complexity and challenges from extensive in vitro characterization to in vivo animal PK studies. General non-clinical safety evaluations of biologics in particular for ADC drugs are outlined including drug-drug interaction (DDI)and metabolite/catabolite assessments. Regulatory guidance on the ADME testing and safety evaluations including immunogenicity as well as bioanalytical considerations are addressed for LMs. In addition, the preclinical and human PK data of two marked ADC drugs (ADCETRIS, as examples are briefly discussed with regard to PK considerations and PK/PD perspectives.

show abstract

“…The pH dependence of the thermal unfolding of H2A-H2B and H3-H4 has been reported, although at lower ionic strengths. 41,42 Between pH 4.5 and 5.5, the mid-point of the thermal unfolding transition, T M , for the H3-H4 heterodimer was [50][51][52][53][54][55][56][57][58][59][60] C, depending on the protein concentration, while the T M of H2A-H2B was 10-15 C lower. Thus, H3-H4 is more stable, but forms fibrils much more readily than H2A-H2B or the individual H2A and H2B monomers which are only partially folded even under stabilizing conditions.…”

Section: Fibrillation Propensity Of Different Histonesmentioning

confidence: 99%

The impact of solubility and electrostatics on fibril formation by the H3 and H4 histones

Topping

Gloss

2011

Protein Science

View full text Add to dashboard Cite

The goal of this study was to examine fibril formation by the heterodimeric eukaryotic histones (H2A-H2B and H3-H4) and homodimeric archaeal histones (hMfB and hPyA1). The histone fold dimerization motif is an obligatorily domain-swapped structure comprised of two fused helix:b-loop:helix motifs. Domain swapping has been proposed as a mechanism for the evolution of protein oligomers as well as a means to form precursors in the formation of amyloid-like fibrils. Despite sharing a common fold, the eukaryotic histones of the core nucleosome and archaeal histones fold by kinetic mechanisms of differing complexity with transient population of partially folded monomeric and/or dimeric species. No relationship was apparent between fibrillation propensity and equilibrium stability or population of kinetic intermediates. Only H3 and H4, as isolated monomers and as a heterodimer, readily formed fibrils at room temperature, and this propensity correlates with the significantly lower solubility of these polypeptides. The fibrils were characterized by ThT fluorescence, FTIR, and far-UV CD spectroscopies and electron microscopy. The helical histone fold comprises the protease-resistant core of the fibrils, with little or no protease protection of the poorly structured N-terminal tails. The highly charged tails inhibit fibrillation through electrostatic repulsion. Kinetic studies indicate that H3 and H4 form a co-fibril, with simultaneous incorporation of both histones. The potential impact of H3 and H4 fibrillation on the cytotoxicity of extracellular histones and a-synuclein-mediated neurotoxicity and fibrillation is considered.

show abstract

Measuring and Increasing Protein Solubility

Cited by 134 publications

References 94 publications

The intrinsic disorder alphabet. III. Dual personality of serine

The intrinsic disorder alphabet. III. Dual personality of serine

An Overall Comparison of Small Molecules and Large Biologics in ADME Testing

The impact of solubility and electrostatics on fibril formation by the H3 and H4 histones

Contact Info

Product

Resources

About