Collaborative study for the establishment of the WHO 3(rd) International Standard for Endotoxin, the Ph. Eur. endotoxin biological reference preparation batch 5 and the USP Reference Standard for Endotoxin Lot H0K354.

An international collaborative study was organised jointly by the World Health Organization (WHO)/National Institute for Biological Standards and Control (NIBSC), the United States Pharmacopeia (USP) and the European Directorate for the Quality of Medicines & HealthCare (EDQM/Council of Europe) for the establishment of harmonised replacement endotoxin standards for these 3 organisations. Thirty-five laboratories worldwide, including Official Medicines Control Laboratories (OMCLs) and manufacturers enrolled in the study. Three candidate preparations (10/178, 10/190 and 10/196) were produced with the same material and same formulation as the current reference standards with the objective of generating a new (3(rd)) International Standard (IS) with the same potency (10 000 IU/vial) as the current (2(nd)) IS, as well as new European Pharmacopoeia (Ph.

Identification of functional regions of Cbp3p, an enzyme-specific chaperone required for the assembly of ubiquinol-cytochrome c reductase in yeast mitochondria.

The Cbp3 protein of Saccharomyces cerevisiae is an enzyme-specific chaperone required for the assembly of ubiquinol-cytochrome c reductase of the mitochondrial respiratory chain. To gain preliminary insight into the role of Cbp3p during assembly, 29 independently isolated mutants were examined to define functional regions of the protein. Mutants were analyzed with respect to respiratory growth, ubiquinol-cytochrome c reductase assembly, and steady state amounts of enzyme subunits and Cbp3p.

Sequence of the Saccharomyces cerevisiae YTP1 gene encoding a deduced novel type-III integral membrane protein with domains of sequence similarity to mitochondrial electron-transport enzymes.

The nucleotide sequence is reported for the Saccharomyces cerevisiae YTP1 (yeast putative transmembrane (TM) protein) gene, encoding a novel deduced protein of 459 amino acids (aa) in length (51 643 Da). The Ytp1 protein appears by computer analysis (hydropathy plots in conjunction with the combined predictions of several Internet on-line programs that deduce protein structure from primary sequence data) to be a type-III integral TM protein containing 10 or 11 TM-spanning domains. Blocks of aa sequence similarity, predominantly to mitochondrial electron transport proteins, are consistent with the notion that Ytp1 is an integral TM protein and may reflect some aspect of its functional role.

Characterization of CBP4, a new gene essential for the expression of ubiquinol-cytochrome c reductase in Saccharomyces cerevisiae.

Nuclear respiratory deficient mutants of Saccharomyces cerevisiae have been screened for lesions in genes affecting ubiquinol-cytochrome c reductase activity. In the present study we describe a new gene, CBP4, whose encoded product is absolutely essential for the activity and expression of the respiratory enzyme. We have cloned and sequenced CBP4, which encodes a 20-kDa protein having no obvious homology to any known protein.