An Arabidopsis mitochondrial proteome project was started for a comprehensive investigation of mitochondrial functions in plants. Mitochondria were prepared from Arabidopsis stems and leaves or from Arabidopsis suspension cell cultures, and the purity of the generated fractions was tested by the resolution of organellar protein complexes applying two-dimensional blue-native/N-[2-hydroxy-1,1-bis...

The profiling of cancer cell secretomes is considered to be a good strategy for identifying cancer-related biomarkers, but few studies have focused on identifying low-molecular-mass (LMr) proteins (<15 kDa) in cancer cell secretomes. Here, we used tricine-SDS-gel-assisted fractionation and LC-MS/MS to systemically identify LMr proteins in the secretomes of five oral cavity squamous cell carcino...

Escherichia coli multidrug resistance protein E (EmrE) is a four transmembrane alpha-helix protein, and a member of the small multidrug resistance protein family that confers resistance to a broad range of quaternary cation compounds (QCC) via proton motive force. The multimeric states of EmrE protein during transport or ligand binding are variable and specific to the conditions of study. To ex...

We have recently developed a kinetically controlled, step-wise affinity cross-linking technique for specific, high-yield, covalent linkage of murine epidermal growth factor (mEGF) via its N terminus to the EGF receptor. EGF receptor from A431 cells was cross-linked to radiolabeled mEGF (125I-mEGF) by this technique and the 125I-mEGF-receptor complex was purified and denatured. Tryptic digestion...

Using the phoA-fusion technology, the recombinant metallothionein (MT) from freshwater crab (Sinopotamon yangtsekiense) has been successfully produced in Escherichia coli. MT purified from the bacterial suspension showed one polypeptide with a molecular weight of 7 kDa by tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Tricine-SDS-PAGE). Western-blotting confirmed the polypep...

Ubiquicidin 29-41 (UBI 29-41) is an antimicrobial peptide with six positively charged residues which preferentially binds to the negatively charged groups present on the microbial membrane by electrostatic interaction at the foci of infection. For this reason UBI 29-41 has been suggested to label by 99m Tc ( 99m Tc-UBI 2941)directly or by using coligand reagents like 6-hydrazinopyridine 3-carbo...

The effect of Tris, Glycine and Tricine on alkaline phosphatase activity was tested at a pH of 8.6, 8.85 and 9.1, using pNPP as substrate. Alkaline phosphatase activity was measured by monitoring the increase in NP absorbance at 420nm using a spectrophotometer. Alkaline phosphatase in Tris buffer at pH 9.1 was found to have the highest activity. MichaelisMenten enzyme kinetic analysis revealed ...