Subject(s)
Bone Marrow/metabolism , Microtubule-Associated Proteins/biosynthesis , Neutropenia/congenital , Adaptor Proteins, Signal Transducing , Adolescent , Apoptosis , Bone Marrow/pathology , Child , Child, Preschool , Disease Progression , Female , Gene Expression Regulation/drug effects , Granulocyte Colony-Stimulating Factor/pharmacology , Granulocyte Colony-Stimulating Factor/therapeutic use , Humans , Inhibitor of Apoptosis Proteins , Lymphoid Enhancer-Binding Factor 1/physiology , Male , Microtubule-Associated Proteins/genetics , Myeloid Cells/metabolism , Neutropenia/drug therapy , Neutropenia/genetics , Neutropenia/metabolism , Neutropenia/pathology , Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics , Precursor Cell Lymphoblastic Leukemia-Lymphoma/metabolism , Precursor Cell Lymphoblastic Leukemia-Lymphoma/pathology , Proteins/genetics , Recombinant Proteins/pharmacology , Recombinant Proteins/therapeutic use , Serine Endopeptidases/genetics , Survivin , Young AdultABSTRACT
Three genes within the genome of E. coli K12 are predicted to encode proteins containing the typical Rieske iron-sulfur cluster-binding motifs. Two of these, hcaC and yeaW, were overexpressed in E. coli BL21 and Tuner (DE3) pLacI. The recombinant proteins were purified and analyzed by UV/Vis- and EPR-spectroscopy. HcaC and YeaW display the typical redox-dependent UV/Vis-spectra of iron-sulfur proteins. The EPR spectrum of reduced HcaC shows characteristic g-values of a Rieske cluster whereas the g-values for YeaW are close to the upper limit for this type of iron-sulfur cluster. Both iron-sulfur clusters could be reduced by dithionite, but not by ascorbate, confirming their classification as low-potential Rieske proteins as derived from the amino acid sequences. A phylogenetic analysis of the two proteins reveals that HcaC clearly segregates with the Rieske ferredoxins of class IIB oxygenases whereas the classification of YeaW remains doubtful.