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Biomedical subjects

Clare Woodward

Publications and source records attributed to Clare Woodward.

8 recordsLinked to original sources
PubMed2005

Assessing positive and negative changes in the aftermath of adversity: psychometric evaluation of the changes in outlook questionnaire.

The Changes in Outlook Questionnaire (CiOQ; S. Joseph, R. Williams, & W. Yule, 1993) is a 26-item self-report measure that was designed to assess positive and negative changes in the aftermath of adversity. This article had 3 aims: 1st, to investigate the factor structure of the CiOQ; 2nd, to test for internal consistency reliability and convergent and discriminant validity; and, 3rd, to investigate the association between positive and negative changes in outlook, posttraumatic stress, and psychological distress. Three studies are reported. Study 1 provides evidence that positive and negative changes are statistically separable and that the 2-factor model is a better fit than the 1-factor model. Studies 2 and 3 provide evidence for internal consistency reliability, convergent and discriminant validity of the CiOQ, and its associations with posttraumatic stress and psychological distress. In conclusion, the CiOQ has much promise for research on responses to stressful and traumatic events.

Adult↗
PubMed2004

Native-like conformations are sampled by partially folded and disordered variants of bovine pancreatic trypsin inhibitor.

Partially folded conformational ensembles of bovine pancreatic trypsin inhibitor (BPTI) are accessed by replacing Cys 5, 30, 51, and 55 by alpha-amino-n-butyric acid (Abu) while retaining the disulfide between Cys 14 and 38; the resultant variant is termed [14-38](Abu). Two new analogues with modifications in the beta-turn, P26D27[14-38](Abu) and N26G27K28[14-38](Abu), are compared to partially folded [14-38](Abu), as well as to [R](Abu), the unfolded protein with all six Cys residues replaced by Abu. Structural features of the new analogues of [14-38](Abu) have been determined by circular dichroism (CD), one-dimensional (1)H NMR, and 8-anilino-1-naphthalenesulfonic acid (ANS) fluorescence experiments. Both analogues are more disordered than the parent [14-38](Abu), but while P26D27[14-38](Abu) has a small population of native-like conformations observed by NMR, no ordered structure is detected for N26G27K28[14-38](Abu). Trypsin inhibition assays were carried out using a modified rat trypsin, C191A/C220A, that minimizes cleavage of unfolded peptides. Both [14-38](Abu) and P26D27[14-38](Abu) significantly inhibit modified trypsin. N26G27K28[14-38](Abu) has low but measurable inhibitor activity, while [R](Abu) has no activity even when in very high molar excess relative to trypsin. ANS fluorescence is enhanced by [14-38](Abu) and by both variants but not by [R](Abu). We conclude that partially folded ensembles of BPTI, even those with little or no CD- or NMR-detectable structure, contain minor populations of native-like conformations. Partially folded [14-38](Abu) and both variants, as well as [R](Abu), have enhanced negative ellipticity in CD spectra acquired in the presence of the osmolyte trimethylamine N-oxide (TMAO). TMAO-induced structure is formed cooperatively, as indicated by thermal unfolding curves. Inhibitor activity as a function of TMAO concentration implies that the osmolyte-induced structure is native-like for [14-38](Abu) and P26D27[14-38](Abu) and is probably native-like for N26G27K28[14-38](Abu). [R](Abu) also shows increased CD-detected structure in the presence of TMAO, but such structure is likely to be collapsed and non-native.

Amino Acid Substitution↗
PubMed2003

Positive change processes and post-traumatic growth in people who have experienced childhood abuse: understanding vehicles of change.

Post-traumatic growth is an emerging area of research concerned with the positive psychological changes that can follow the experience of traumatic events. The aim of this study is to explore themes of post-traumatic growth within personal experience narratives of individuals who have experienced some form of early emotional, physical, or sexual abuse. Using thematic analysis, we identified three domains of themes related to positive change processes: inner drive toward growth, vehicles of change, and psychological changes. Understanding the different vehicles of change has implications for facilitating post-traumatic growth in clients who have experienced traumatic events.

Adult↗
PubMed2002

Hydrogen exchange, core modules, and new designed proteins.

A strategy for design of new proteins that mimic folding properties of native proteins is based on peptides modeled on the slow exchange cores of natural proteins. We have synthesized peptides, called core modules, that correspond to the elements of secondary structure that carry the very slowest exchanging amides in a protein. The expectation is that, if soluble in water, core modules will form conformational ensembles that favor native-like structure. Core modules modeled on natural bovine pancreatic trypsin inhibitor have been shown by NMR studies to meet this expectation. The next step toward production of a native state mimic is to further shift the conformational bias of a core module toward more ordered structure by promoting module-module interactions that are mutually stabilizing. For this, two core modules were incorporated into a single molecule by means of a long cross-link. From a panel of several two-module peptides, one very promising lead emerged; it is called BetaCore. BetaCore is monomeric in water and forms a new fold composed of a four-stranded, antiparallel beta-sheet. The single, dominant conformation of BetaCore is characterized by various NMR experiments. Here we compare the individual core module to the two-module BetaCore and discuss the progressive stabilization of intramodule structure and the formation of new intermodule interactions.

Amino Acid Sequence↗
PubMed2002

A highly destabilizing mutation, G37A, of the bovine pancreatic trypsin inhibitor retains the average native conformation but greatly increases local flexibility.

A point mutation, G37A, on the surface of bovine pancreatic trypsin inhibitor (BPTI) destabilizes the protein by approximately 5 kcal/mol, which is very high for addition of one methyl group. In wild-type (WT) BPTI, Gly 37 HN is in an unusual NH-aromatic-NH network of interactions with the ring of Tyr 35 and the side chain HN of Asn 44. G37A was designed to disrupt this interaction, since the phi and psi backbone angles of G37 are not favorable for an amino acid containing a beta-carbon. Investigations of the structure and dynamics by NMR methods show that G37A retains the average WT structure. The NH-aromatic-NH interactions remain intact, as indicated by NOEs and the large upfield ring current shift (approximately 4 ppm) of A37 HN. The NMR structure, confirmed by molecular modeling calculations, requires phi and psi backbone angles that are highly destabilizing when alanine is in position 37. Although the average structure is essentially unchanged, the dynamics are altered dramatically. Many residues in the region of the mutation have increased flexibility, as probed by aromatic ring flip rates and native state hydrogen exchange. We conclude that a large fraction of the destabilization arises from maintaining A37 in a high-energy conformation. This suggests that disruption of the NH-aromatic-NH network is energetically very costly, and may involve other cooperatively linked interactions. The results illustrate the importance of the Gly-Gly sequence at positions 36 and 37 and the 37 HN-35 aromatic interaction to the stability, folding, and dynamics of the BPTI.

Alanine↗
PubMed2002

Native-like interactions favored in the unfolded bovine pancreatic trypsin inhibitor have different roles in folding.

Folding kinetics of a series of bovine pancreatic trypsin inhibitor (BPTI) variants with similar stabilities and structures have been measured. All are strongly destabilized relative to WT. In Y21A, F22A, Y23A, G37A, and F45A, the three native disulfide bonds are retained. In RM(14-38), Cys14 and Cys38 thiols are methylated while C30-C51 and C5-C55 disulfides remain intact. At pH 2 and 20 degrees C, relaxation rate constants of the major kinetic phase range from approximately 10 ms to 0.71 s in the absence of denaturant. All mutants except G37A exhibit standard two-state behavior. Y21A, F22A, and Y23A fold much more slowly than other mutants. The experiments were designed to test the hypothesis that native-like structure detected in the unfolded BPTI is important in folding. Two native-like contacts are implied by NOEs in reduced and unfolded BPTI, between residues Tyr23 and Ala25, and between Gly37 NH and the Tyr35 ring. The results support an earlier hypothesis that formation of the central beta-hairpin, monitored by a local native interaction between Tyr23 and Ala25, is crucial to initiation of BPTI folding. The second native-like contact is important, not in folding initiation, but in preventing a kinetic trap later in the process. Evidence for this comes from mutant G37A, which behaves very differently from the others in displaying a phenomenon called rollover. G37A is, to our knowledge, the first reported case in which a single-site replacement causes rollover, while the wild type and all other known mutants of the same protein show typical two-state chevron plots. The best explanation is that the G37A mutation introduces a kinetic trap of the type described by Chan and Dill [(1998) Proteins 30, 2-33]. In native BPTI, there is an unusual polar interaction between the ring of Tyr35 and the backbone NH of Gly37. Our results suggest that the NH-aromatic interaction between residues 37 and 35 is important throughout folding in stabilizing native-like loop conformations and in preventing the flexible loops from being trapped in nonfunctional conformations during later stages of folding.

Animals↗
PubMed2002

BetaCore, a designed water soluble four-stranded antiparallel beta-sheet protein.

BetaCore is a designed approximately 50-residue protein in which two BPTI-derived core modules, CM I and CM II, are connected by a 22-atom cross-link. At low temperature and pH 3, homo- and heteronuclear NMR data report a dominant folded ('f') conformation with well-dispersed chemical shifts, i, i+1 periodicity, numerous long-range NOEs, and slowed amide hydrogen isotope exchange patterns that is a four-stranded antiparallel beta-sheet with nonsymmetrical and specific association of CM I and CM II. BetaCore 'f' conformations undergo reversible, global, moderately cooperative, non-two-state thermal transitions to an equilibrium ensemble of unfolded 'u' conformations. There is a significant energy barrier between 'f' and 'u' conformations. This is the first designed four-stranded antiparallel beta-sheet that folds in water.

Amino Acid Sequence↗