48 research outputs found
Identificazione di un nuovo inibitore di serin-proteasi (68 kDa) nell’encefalo di ratto.
Pig kidney dopa decarboxylase. Structure and function
Pig kidney dopa decarboxylase. Structure and functio
SUBSTITUTION OF ARGININE FOR LYSINE 134 ALTERS ELECTROSTATIC PARAMETERS OF THE ACTIVE-SITE IN SHARK CU,ZN SUPEROXIDE-DISMUTASE RID D-1236-2011
Protein minimization: characterization of the synthetic cyclic dodecapeptide corresponding to the reactive site region of the oil rape trypsin inhibitor type-III
The design of minimal units required for enzyme inhibition is a major field of interest in structural biology and biotechnology. The successful design of the cyclic dodecapeptide corresponding to the Phe17–Val28 reactive site amino acid sequence of the lowmolecular-mass trypsin inhibitor RTI-III from Brassica napus (micro-RTI-III) and of the recombinant murine dihydrofolate reductase-(DHFR-)micro-RTI-III fusion protein (DHFR-micro-RTI-III) is reported here. Micro-RTI-III was synthesized using a stepwise solid-phase approach based on the standard Fmoc chemistry, purified by RP-HPLC, and oxidatively refolded. DHFRmicro- RTI-III was expressed in Escherichia coli, purified bymetal-chelate affinity chromatography , and oxidatively refolded. The affinityof micro-RTI-III for bovine trypsin (Kd ¼ 1:6 109 M) is similar to that determined for DHFR-micro-RTI-III (Kd ¼ 6:3 1010 M) and native RTI-III (Kd ¼ 2:9 1010 M), at pH 8.2 and 22.0 C. Remarkably, micro-RTI-III protects the DHFR domain of DHFR-micro-RTI-III from trypsin digestion. Micro-RTI-III is a new minimal trypsin inhibitor and may be regarded as a tool in protein structure–function studies and for developing multifunctional and multidomain proteinase inhibitors
Accuracy of an immune diagnostic assay based on RD1 selected epitopes for active tuberculosis in a clinical setting: a pilot study
A previous case-control study reported that an in-vitro interferon (IFN)-gamma response to early secreted antigenic target (ESAT)-6 selected peptides was associated with active tuberculosis (A-TB). The objective of the present pilot study was to evaluate the diagnostic accuracy of this assay for TB disease in a clinical setting. An IFN-gamma ELISPOT assay was performed on samples from patients with suspected A-TB using two peptides selected from ESAT-6 protein and three peptides selected from culture filtrate 10 (CFP-10) proteins. The results were compared with those obtained by two commercially available assays approved for diagnosis of TB infection (T SPOT-TB and QuantiFERON-TB Gold) which use ESAT-6/CFP-10 (RD1) overlapping peptides. Sensitivity to the RD1 selected peptides was 70% (positive for 16 of 23 patients with microbiologically diagnosed A-TB) and specificity was 91% (positive for three of 32 controls). In contrast, the sensitivity and specificity were 91% and 59%, respectively, for T SPOT-TB, and were 83% and 59%, respectively, for QuantiFERON-TB Gold. The RD1 selected peptides assay had the highest diagnostic odds ratio for A-TB. Thus, the results suggest that an assay based on RD1 selected peptides has a higher diagnostic accuracy for A-TB in a clinical setting compared with commercially available assays based on RD1 overlapping peptides