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by partial or incorrect folding, probably due to the reducing environment of theE. coli cytoplasm. However, it was soon realised ...

Table 8.4 Summary of chromatographic techniques commonly used in protein purification Technique Property exploited Capacity Reso ...

(ligand) is bound to an insoluble support. When a crude mixture of proteins contain- ing the protein of interest is passed throu ...

Reduction of the intracellular concentration of toxic proteins: Some cloned proteins are toxic to the cell in which they are pr ...

N-Asp-Asp-Asp-Lys-C. Using this approach, granulocyte-macrophage colony-stimulating factor (GMCSF) was cloned in and secreted fr ...

estimate of size and these methods are sufficient. The third method, mass spectrometry, requires expensive specialist instrument ...

the loss of N-terminal lysine was 20 072.8. Clearly mass spectrometry has the ability to provide highly accurate molecular mass ...

To search for sequence homology using computerised databases in order to identify the function of the protein. For example, the ...

introduction of fast atom bombardment (FAB) MS allowed the analysis of large, charged molecules such as proteins and peptides to ...

trypsin) is separated by MS. The ion corresponding to one peptide is selected in the first analyser and collided with argon gas ...

sequence that contained spans of 20–28 amino acid residues, each of which contained predominantly hydrophobic residues flanked o ...

30% of consensus sequences for N-linked attachments are occupied by polysacchar- ide; the nature of the secondary structure at t ...

The first question to be asked about a purified protein is ‘Is it a glycoprotein?’ Glycoprotein bands in gels (e.g. on SDS-polya ...

attempted using as homogeneous a preparation as possible, such preparations having a greater chance of yielding crystals than ma ...

Unfortunately the diffraction pattern alone is insufficient to determine the crystal structure. Each diffraction maximum has bo ...

auxotroph, and then grow this strain in the presence of selenomethionine (a selenium- containing analogue of methionine). Seleno ...

8.5 Proteomics and protein function In order to completely understand how a cell works, it is necessary to understand the functi ...

of a cell is referred to as the genome, and the study of the structure and function of this DNA is called genomics. By analogy, ...

observe the response to extracellular stimuli such as hormones or cytokines; compare pathogenic and non-pathogenic bacterial st ...

where one of the peptides separated for mass-fingerprinting is further fragmented in a second analyser, and from the fragmentati ...