Catalase HPII from Escherichia coli exhibits enhanced resistance to denaturation.

Catalase HPII from Escherichia coli is a homotetramer of 753 residue subunits. The multimer displays a number of unusual structural features, including interwoven subunits and a covalent bond between Tyr415 and His392, that would contribute to its rigidity and stability. As the temperature of a solution of HPII in 50 mM potassium phosphate buffer (pH 7) is raised from 50 to 92 degrees C, the enzyme begins to lose activity at 78 degrees C and 50% inactivation has occurred at 83 degrees C. The inactivation is accompanied by absorbance changes at 280 and 407 nm and by changes in the CD spectrum consistent with small changes in secondary structure. The subunits in the dimer structure remain associated at 95 degrees C and show a significant level of dissociation only at 100 degrees C. The exceptional stability of the dimer association is consistent with the interwoven nature of the subunits and provides an explanation for the resistance to inactivation of the enzyme. For comparison, catalase-peroxidase HPI of E. coli and bovine liver catalase are 50% inactivated at 53 and 56 degrees C, respectively. In 5.6 M urea, HPII exhibits a coincidence of inactivation, CD spectral change, and dissociation of the dimer structure with a midpoint of 65 degrees C. The inactive mutant variants of HPII which fold poorly during synthesis and which lack the Tyr-His covalent bond undergo spectral changes in the 78 to 84 degrees C range, revealing that the extra covalent linkage is not important in the enhanced resistance to denaturation and that problems in the folding pathway do not affect the ultimate stability of the folded structure.

Lifestyle in Curacao: smoking, alcohol consumption, eating habits and exercise

The Curacao Health Study was carried out among a randomized sample (n = 2248, response rate = 85 percent) of the adult non-institutionalized population in order to assess aspects of lifestyle that may pose health risks. Factors examined were tobacco and alcohol use, eating habits and exercise behaviour. Outcome variables were cross-tabulated by gender, age and socioeconomic status. 17.1 percent of the participants were smokers and 20.5 percent regular drinkers, including 6.3 percent of the men who consumed alcohol excessively (4 or more glasses of alcohol a day). 75 percent of the participants did not excercise regularly, 37 percent did not eat vegetables daily, and half did not eat fruit daily. Other poor eating habits were the addition of extra sugar and salt to prepared food by 33 percent and 20 percent of the participants, respectively. On the whole, men had less healthy lifestyles than women, with the exception of execise behaviour. People of high socioeconomic status (SES) drank less alcohol, and exercised more often than those of low SES. Considering the high prevalence of diabetes mellitus and hypertension in the Caribbean, research of lifestyle factors in other Caribbean countries is required to facilitate the development of regional prevention and intervention programmes.