Electron tunneling in proteins: coupling through a beta strand.

Electron coupling through a beta strand has been investigated by measurement of the intramolecular electron-transfer (ET) rates in ruthenium-modified derivatives of the beta barrel blue copper protein Pseudomonas aeruginosa azurin. Surface histidines, introduced on the methionine-121 beta strand by mutagenesis, were modified with a Ru(2,2'-bipyridine)2(imidazole)2+ complex. The Cu+ to Ru3+ rate constants yielded a distance-decay constant of 1.1 per angstrom, a value close to the distance-decay constant of 1.0 per angstrom predicted for electron tunneling through an idealized beta strand. Activationless ET rate constants in combination with a tunneling-pathway analysis of the structures of azurin and cytochrome c confirm that there is a generally efficient network for coupling the internal (native) redox center to the surface of both proteins.

Spontaneous pneumothorax associated with ankylosing spondylitis.

OBJECTIVE: To review the incidence and clinical characteristics of spontaneous pneumothorax in patients with ankylosing spondylitis (AS). METHODS: This is a retrospective observational cohort study. Chest radiographs and medical records of patients with a diagnosis of AS from 1993 to 2003 in a tertiary referral centre were reviewed. RESULTS: A total of 1028 patients with a diagnosis of AS were identified from July 1993 to July 2003. Twenty-two patients had typical apical lung fibrotic changes in the chest radiographs (22/1028, 2.1%). Three of these patients (3/22, 13.6%) with lung disease had pneumothorax. Two patients had recurrences and received video-assisted thoracoscopic surgery (VATS) on the second attack. The third patient received talc pleurodesis on the first attack and did not have a recurrence. The incidence of spontaneous pneumothorax in patients with AS was 0.29% (3/1028) with an incidence density of 64.85/100,000 patient-yr (95% confidence interval: 66.17-63.57/100,000). CONCLUSION: Spontaneous pneumothorax appears to be exceedingly rare in AS unless there is an underlying fibrocystic lung disease, in which case its occurrence is not uncommon. Cigarette smoking may be an important co-factor. Once developed, recurrences are common even after treatment. Prophylactic procedures should thus be considered for the first attack of spontaneous pneumothorax.

Seroprevalence of SARS coronavirus antibody in household contacts.

Between March and July 2003, 671 cases of severe acute respiratory syndrome (SARS) were diagnosed in Taiwan with a total of 84 fatalities. After the epidemic, a serological survey was conducted involving the asymptomatic household contacts. Household contacts of 13 index patients were enrolled in the study. Contact history and clinical symptoms of the household contacts were recorded by standardized questionnaires. Blood samples of patients and household contacts were collected at least 28 days after symptom onset in the index patients or household exposure in the contacts for SARS-associated coronavirus (SARS-CoV) IgG testing. On the basis of this investigation, 29 persons (25 adults and 4 children) were identified as having had unprotected exposure to the index cases before infection-control practices were implemented. Laboratory evaluation of clinical specimens showed no evidence of transmission of SARS-CoV infection to any contacts. This investigation demonstrated that subclinical transmission among household contacts was low in the described setting.

Identification of chemical modification sites on metalloproteins by capillary electrophoresis.

Capillary electrophoresis (CE) has been used to separate the peptides obtained from tryptic digestion of ruthenium-modified cytochrome c. The modified peptide was identified from a comparison of the elution profile and absorbance characteristics of the native and modified proteins. Automatic fraction collection on the CE instrument provides sufficient amounts of this modified peptide for amino acid sequencing. Capillary electrophoresis has also been used to monitor the modification reaction and to optimize the modification efficiency. The methodologies have been extended to myoglobin in order to monitor the modification of multiple surface sites.

Electron transfer in ruthenium-modified proteins.

Photochemical techniques have been used to measure the kinetics of intramolecular electron transfer in Ru(bpy)2(im)(His)2(+)-modified (bpy = 2,2'-bipyridine; im = imidazole) cytochrome c and azurin. A driving-force study with the His33 derivatives of cytochrome c indicates that the reorganization energy (lambda) for Fe2+-->Ru3+ ET reactions is 0.8 eV. Reductions of the ferriheme by either an excited complex, *Ru2+, or a reduced complex, Ru+, are anomalously fast and may involve formation of an electronically excited ferroheme. The distance dependence of Fe2+-->Ru3+ and Cu+-->Ru3+ electron transfer in 12 different Ru-modified cytochromes and azurins has been analyzed using a tunneling-pathway model. The ET rates in 10 of the 12 systems exhibit an exponential dependence on metal-metal separation (decay constant of 1.06 A-1) that is consistent with prediction of the pathway model.