Dataset Information

Exploring local solvation environments of a heme protein using the spectroscopic reporter 4-cyano-l-phenylalanine.

ABSTRACT: The vibrational reporter unnatural amino acid (UAA) 4-cyano-l-phenylalanine (pCNF) was genetically incorporated individually at three sites (5, 36, and 78) in the heme protein Caldanaerobacter subterraneus H-NOX to probe local hydration environments. The UAA pCNF was incorporated site-specifically using an engineered, orthogonal tRNA synthetase in E. coli. The ability of all of the pCNF-containing H-NOX proteins to form the ferrous CO, NO, or O2 ligated and unligated states was confirmed with UV-Vis spectroscopy. The solvation state at each site of the three sites of pCNF incorporation was assessed using temperature-dependent infrared spectroscopy. Specifically, the frequency-temperature line slope (FTLS) method was utilized to show that the nitrile group at site 36 was fully solvated and the nitrile group at site 78 was de-solvated (buried) in the heme pocket. The nitrile group at site 5 was found to be partially solvated suggesting that the nitrile group was involved in moderate strength hydrogen bonds. These results were confirmed by the determination of the X-ray crystal structure of the H-NOX protein construct containing pCNF at site 5.

SUBMITTER: Kearney C

PROVIDER: S-EPMC5944249 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature

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Exploring local solvation environments of a heme protein using the spectroscopic reporter 4-cyano-l-phenylalanine.

Kearney Caroline C Olenginski Lukasz T LT Hirn Trexler D TD Fowler Gwendolyn D GD Tariq Daniyal D Brewer Scott H SH Phillips-Piro Christine M CM

RSC advances 20180410 24

The vibrational reporter unnatural amino acid (UAA) 4-cyano-l-phenylalanine (pCNF) was genetically incorporated individually at three sites (5, 36, and 78) in the heme protein Caldanaerobacter subterraneus H-NOX to probe local hydration environments. The UAA pCNF was incorporated site-specifically using an engineered, orthogonal tRNA synthetase in E. coli. The ability of all of the pCNF-containing H-NOX proteins to form the ferrous CO, NO, or O2 ligated and unligated sta ...[more]

PMID: 29780583

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Project description:IntroductionObesity is a global pandemic that affects all socioeconomic strata, however, the highest figures have been observed in the most disadvantaged social groups. Evidence from the USA and Canada showed that specific urban settings encourage obesogenic behaviour in the population living and/or working there. We aim to examine the evidence on the association between local food environments and obesity in the UK, Ireland, Australia and New Zealand.MethodsSix databases from 1990 to 2017 will be searched: MEDLINE (Ovid), Embase (Ovid), Scopus, The Cumulative Index to Nursing and Allied Health Literature (CINAHL), Applied Social Sciences Index and Abstracts (ASSIA) and Web of Science. Grey literature will also be sought by searching Opengrey Europe, The Grey Literature Report and relevant government websites. Additional studies will be retrieved from the reference lists of the selected articles. It will include cohort, longitudinal, case study and cross-sectional studies that have assessed the relationship between local food environments and obesity in the UK, Ireland, Australia and New Zealand regardless of sex, age and ethnicity of the population. Two researchers will independently select the studies and extract the data. Data items will incorporate: author names, title, study design, year of study, year exposure data collected, country, city, urban/rural, age range, study exclusions, special characteristics of study populations, aims, working definitions of food environments and food outlets, exposure and methods of data collection, outcomes and key findings. A narrative synthesis and a summary of the results will be produced separately for children and adults, according to the type of food exposure-outcome. All the selected studies will be assessed using The Quality Assessment Tool for Observational Cohort and Cross-Sectional Studies.Ethics and disseminationThis study will be based on published literature, and therefore ethical approval has not been sought. Our findings will be presented at relevant national and international scientific conferences and published in a peer-reviewed journal.

Dataset Information

Exploring local solvation environments of a heme protein using the spectroscopic reporter 4-cyano-l-phenylalanine.

Publications

Exploring local solvation environments of a heme protein using the spectroscopic reporter 4-cyano-l-phenylalanine.

Similar Datasets

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