A5024971805- Protein Structure and Dynamics
- Enzyme Structure and Function
- Advanced NMR Techniques and Applications
- Molecular spectroscopy and chirality
- Heat shock proteins research
- Photosynthetic Processes and Mechanisms
- NMR spectroscopy and applications
- Chemical Synthesis and Analysis
- Monoclonal and Polyclonal Antibodies Research
- Biochemical and Structural Characterization
- Viral Infections and Outbreaks Research
- Alzheimer's disease research and treatments
- NF-κB Signaling Pathways
- Endoplasmic Reticulum Stress and Disease
- Amyloidosis: Diagnosis, Treatment, Outcomes
- Toxin Mechanisms and Immunotoxins
- Viral Infections and Vectors
- RNA and protein synthesis mechanisms
- Bacillus and Francisella bacterial research
- Glycosylation and Glycoproteins Research
- Metabolism and Genetic Disorders
- Immune Response and Inflammation
- Bacterial Genetics and Biotechnology
- Photoreceptor and optogenetics research
- Mass Spectrometry Techniques and Applications
Scripps Research Institute
2013-2024
Cornell University
2004
University of California, San Francisco
2001
University of Groningen
1993-1997
Graduate School Experimental Plant Sciences
1991
Random coil chemical shifts are commonly used to detect secondary structure elements in proteins shift index calculations. While this technique is very reliable for folded proteins, application unfolded reveals significant deviations from measured random certain nuclei. some of these can be ascribed residual the protein, others clearly caused by local sequence effects. In particular, amide nitrogen, proton, and carbonyl carbon highly sensitive amino acid sequence. We present a detailed,...
Human amylin, or islet amyloid polypeptide, is a peptide cosecreted with insulin by the beta cells of pancreatic islets Langerhans. The 37-residue, C-terminally amidated human amylin derives from proprotein that undergoes disulfide bond formation in endoplasmic reticulum and then subjected to four enzymatic processing events immature secretory granule. forms both intracellular extracellular deposits pancreas most type II diabetic subjects, likely reflecting compromised cell function. In...
A template-assisted conformational change of the cellular prion protein (PrPC) from a predominantly helical structure to an amyloid-type with higher proportion β-sheet is thought be causative factor in diseases. Since flexibility polypeptide likely contribute ability PrPC undergo that leads infective state, we have undertaken comprehensive examination dynamics two recombinant Syrian hamster PrP fragments, PrP(29−231) and PrP(90−231), using 15N NMR relaxation measurements. The molecular...
Beta-turns are common conformations that enable proteins to adopt globular structures, and their formation is often rate limiting for folding. Beta-turn mimics, molecules replace the i + 1 2 amino acid residues of a beta-turn, envisioned act as folding nucleators by preorganizing pendant polypeptide chains, thereby lowering activation barrier beta-sheet formation. However, crucial kinetic experiments demonstrate beta-turn mimics can strong in context cooperatively protein have not been...
Filoviruses, including Marburg virus (MARV) and Ebola (EBOV), cause fatal hemorrhagic fever in humans non-human primates. All filoviruses encode a unique multi-functional protein termed VP35. The C-terminal double-stranded (ds)RNA-binding domain (RBD) of VP35 has been implicated interferon antagonism immune evasion. Crystal structures the RBD from two ebolaviruses have previously demonstrated that viral caps ends dsRNA. However, it is not yet understood how expanses dsRNA backbone, between...
Cows produce antibodies with a disulfide-bonded antigen-binding domain embedded within ultralong heavy chain third complementarity determining regions. This "knob" is analogous to natural cysteine-rich peptides such as knottins in that it small and stable but can accommodate diverse loops disulfide bonding patterns. We immunized cattle SARS-CoV-2 spike found CDR H3 could neutralize several viral variants at picomolar IC
The co-chaperone p23 forms a complex with the chaperone Hsp90 that mediates folding pathway leading to production of functional steroid receptors. Solution NMR spectroscopy has been used characterize sites interaction between and p23. Titration results in selective broadening certain cross-peaks 15N-1H heteronuclear single quantum correlation (HSQC) spectrum. on have localized by dissection into single-domain two-domain constructs. N-terminal (N) domain does not affect spectrum either...
Significance Molecular chaperones play key roles in maintaining protein homeostasis within cells. Membrane face particular challenges, as they not only protect highly aggregation-prone membrane substrates, but also need to achieve tight spatiotemporal coordination of their chaperone cycle. In this work, biochemical and NMR analyses address these questions for the first time, our knowledge, define complete cycle cpSRP43, an ATP-independent dedicated integral proteins. The study reveals that...
Significance Stress-response transcription factors turn on hundreds of genes, and their activity must be turned off completely quickly. The inhibitor protein IκBα turns NFκB by forming a transient ternary complex with the NFκB–DNA then promoting DNA dissociation (molecular stripping). Here we report mutant that is impaired in its ability to strip from DNA. forms more stable complex, biophysical characterization shows what looks like “in act stripping.” We also show single-cell nuclear export...
The cyclic AMP response element (CRE) binding protein (CREB) is a transcription factor that contains 280-residue N-terminal transactivation domain and basic leucine zipper mediates interaction with DNA. comprises three subdomains, the glutamine-rich domains Q1 Q2 kinase inducible activation (KID). NMR chemical shifts show isolated subdomains are intrinsically disordered but have propensity to populate local elements of secondary structure. exhibit for formation short β-hairpin motifs...
The region of the surface histidine‐containing protein (HPr) which interacts with A domain mannitol‐specific Enzyme II (IIA mtl ) has been mapped by titrating A‐domain into a solution 15 N‐labeled HPr and monitoring effects on amide proton nitrogen chemical shifts via heteronuclear single quantum correlation spectroscopy (HSQC). Fourteen eighty‐five amino acid residues show large changes in either N or 1 H both as result presence IIA while further seventeen experience lesser shifts. Most...
The N-terminal cysteine-rich somatomedin B (SMB) domain (residues 1−44) of the human glycoprotein vitronectin contains high-affinity binding sites for plasminogen activator inhibitor-1 (PAI-1) and urokinase receptor (uPAR). We previously showed that eight cysteine residues recombinant SMB (rSMB) are organized into four disulfide bonds in a linear uncrossed pattern (Cys5−Cys9, Cys19−Cys21, Cys25−Cys31, Cys32−Cys39). In present study, we use an alternative method to show this bond arrangement...
We present a detailed investigation of unfolded and partially folded states mutant apomyoglobin (apoMb) where the distal histidine has been replaced by phenylalanine (H64F). Previous studies have shown that substitution His64, located in E helix native protein, stabilizes equilibrium molten globule leads to an increase folding rate change pathway. Analysis changes chemical shift backbone flexibility, detected via [1H]-15N heteronuclear nuclear Overhauser effect measurements, indicates only...
Abstract The aggregation pathway of transthyretin (TTR) proceeds through rate-limiting dissociation the tetramer and partial misfolding monomers, which assemble into amyloid structures a downhill polymerization mechanism. structural features aggregation-prone monomeric intermediate are poorly understood. Characterization amyloidogenic intermediates is challenging due to their propensity aggregate at concentrations necessary for studies. NMR relaxation dispersion offers unique opportunity...