Spectroscopic studies reveal details of substrate-induced conformational changes distant from the active site in isopenicillin N synthase.

Isopenicillin N synthase (IPNS) catalyzes formation of the ß-lactam and thiazolidine rings of isopenicillin N from its linear tripeptide l-δ-(α-aminoadipoyl)-l-cysteinyl-d-valine (ACV) substrate in an iron- and dioxygen (O2)-dependent four-electron oxidation without precedent in current synthetic chemistry. Recent X-ray free-electron laser studies including time-resolved serial femtosecond crystallography show that binding of O2 to the IPNS-Fe(II)-ACV complex induces unexpected conformational changes in α-helices on the surface of IPNS, in particular in α3 and α10. However, how substrate binding leads to conformational changes away from the active site is unknown. Here, using detailed 19F NMR and electron paramagnetic resonance experiments with labeled IPNS variants, we investigated motions in α3 and α10 induced by binding of ferrous iron, ACV, and the O2 analog nitric oxide, using the less mobile α6 for comparison. 19F NMR studies were carried out on singly and doubly labeled α3, α6, and α10 variants at different temperatures. In addition, double electron-electron resonance electron paramagnetic resonance analysis was carried out on doubly spin-labeled variants. The combined spectroscopic and crystallographic results reveal that substantial conformational changes in regions of IPNS including α3 and α10 are induced by binding of ACV and nitric oxide. Since IPNS is a member of the structural superfamily of 2-oxoglutarate-dependent oxygenases and related enzymes, related conformational changes may be of general importance in nonheme oxygenase catalysis.

Age and gender correlation of gonial angle, ramus height and bigonial width in dentate subjects in a dental school in Far North Queensland.

BACKGROUND: This study aimed to determine if mandibular parameters (gonial angle, bigonial width and ramus height) measured from panoramic radiographs, can be used to determine a correlation with an individual's age and gender in dentate subjects in Far North Queensland. MATERIAL AND METHODS: The study utilised 2699 randomly selected panoramic radiographs of patients between the ages of 19-69 years, from which 220 fulfilled the inclusion criteria. Each panoramic radiograph was analysed and the above three parameters recorded and measured. These values were collated into appropriate age and gender groups and subjected to statistical analysis. RESULTS: The mean age of the participants was 44.1±14.41, with males being shown to have a statistically significant larger ramus height and bigonial width than females (P<0.0001 for both). Females, on the other hand, were shown to have a significantly larger gonial angle than males (P<0.0002). General trends revealed gonial angle to increase with age, whilst bigonial width and ramus height were shown to decrease with age. CONCLUSIONS: The assessment of mandibular morphology through radiographic measurements may be useful in estimating an individual's age and gender when comparing to a known population standard. KEY WORDS: Bigonial width, gonial angle, panoramic radiograph, ramus height.