Structure and denaturation of 4-chlorobenzoyl coenzyme A dehalogenase from Arthrobacter sp. strain TM-1

The secondary structure of the trimeric protein 4-chlorobenzoyl coenzyme A dehalogenase from Arthrobacter sp. strain TM-1, the second of three enzymes involved in the dechlorination of 4-chlorobenzoate to form 4-hydroxybenzoate, has been examined. E(mM) for the enzyme was 12.59. Analysis by circular dichroism spectrometry in the far uv indicated that 4-chlorobenzoyl coenzyme A dehalogenase was composed mostly of alpha-helix (56%) with lesser amounts of random coil (21%), beta-turn (13%) and beta-sheet (9%). These data are in close agreement with a computational prediction of secondary structure from the primary amino acid sequence, which indicated 55.8% alpha-helix, 33.7% random coil and 10.5% beta-sheet; the enzyme is, therefore, similar to the 4-chlorobenzoyl coenzyme A dehalogenase from Pseudomonas sp. CBS-3. The three-dimensional structure, including that of the presumed active site, predicted by computational analysis, is also closely similar to that of the Pseudomonas dehalogenase. Study of the stability and physicochemical properties revealed that at room temperature, the enzyme was stable for 24 h but was completely inactivated by heating to 60 degrees C for 5 min; thereafter by cooling at 1 degrees C min(-1) to 45 degrees C, 20.6% of the activity could be recovered. Mildly acidic (pH 5.2) or alkaline (pH 10.1) conditions caused complete inactivation, but activity was fully recovered on returning the enzyme to pH 7.4. Circular dichroism studies also indicated that secondary structure was little altered by heating to 60 degrees C, or by changing the pH from 7.4 to 6.0 or 9.2. Complete, irreversible destruction of, and maximal decrease in the fluorescence yield of the protein at 330-350 nm were brought about by 4.5 M urea or 1.1 M guanidinium chloride. Evidence was obtained to support the hypothetical three-dimensional model, that residues W140 and W167 are buried in a non-polar environment, whereas W182 appears at or close to the surface of the protein. At least one of the enzymes of the dehalogenase system (the combined 4-chlorobenzoate:CoA ligase, the dehalogenase and 4-hydroxybenzoyl coenzyme A thioesterase) appears to be capable of association with the cell membrane.

[Conference report] Magic and mathematics: The life and work of John Dee. National Maritime Museum, Greenwich, 13-14 June 2003

Tony Mann provides a report of a two-day meeting "Magic and mathematics: The life and work of John Dee" held from 13-14 June 2003 at the National Maritime Museum, Greenwich.

[Recording review] William Herschel, Symphonies Nos. 2, 8, 12, 13, 14, 17

Tony Mann provides a review of the recording: William Herschel, Symphonies Nos. 2, 8, 12, 13, 14, 17, soloists, London Mozart Players / Matthias Bamert, Chandos 10048, 2003, (68 mins). [CD]

‘The trouble with Year 10’: 13 – 16 year old school students’ attitudes to higher education

This study looked at the impact of Widening Participation interventions on the attitudes of young people towards higher education. A total of 2731 adolescents aged 13–16 years completed a self-report measure of their attitudes to higher education, general and academic self concept and identification with school, family and peers. This was matched with data on the students’ academic attainment and social backgrounds. As expected, attainment scores were significantly positively correlated with take up of Widening Participation activities aimed at increasing participation in higher education, attitudes towards going to university and academic motivation. However, attainment was negatively correlated with perceptions of family attending university and identification with family. Regression analyses found that perceptions of family views about attending university were not a predictor of taking part in Widening Participation activities but were a predictor of attitudes towards higher education. Students in Year 10 aged 14–15 were significantly more negative on most factors than either older or younger students.