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Recombinant proteins are often expressed in the form of insoluble inclusion bodies in bacteria. To facilitate refolding of recombinant proteins obtained from inclusion bodies, 0.1 to 1 M arginine is customarily included in solvents used for refolding the proteins by dialysis or dilution. In addition, arginine at higher concentrations, e.g., 0.5-2 M, can be used to extract active, folded proteins from insoluble pellets obtained after lysing Escherichia coli cells. Moreover, arginine increases the yield of proteins secreted to the periplasm, enhances elution of antibodies from Protein-A columns, and stabilizes proteins during storage. All these arginine effects are apparently due to suppression of protein aggregation. Little is known, however, about the mechanism. Various effects of solvent additives on proteins have been attributed to their preferential interaction with the protein, effects on surface tension, or effects on amino acid solubility. The suppression of protein aggregation by arginine cannot be readily explained by either surface tension effects or preferential interactions. In this review we show that interactions between the guanidinium group of arginine and tryptophan side chains may be responsible for suppression of protein aggregation by arginine.

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The crystal structure of a microbial transglutaminase from Streptoverticillium mobaraense has been determined at 2.4 Å resolution. The protein folds into a platelike shape, and has one deep cleft at the edge of the molecule. Its overall structure is completely different from that of the factor XIII-like transglutaminase, which possesses a cysteine protease-like catalytic triad. superimpose well on the catalytic triad "Cys-HisAsp" of the factor XIII-like transglutaminase, in this order. The secondary structure frameworks around these residues are also similar to each other. These results imply that both transglutaminases are related by convergent evolution; however, the microbial transglutaminase has developed a novel catalytic mechanism specialized for the cross-linking reaction. The structure accounts well for the catalytic mechanism, in which Asp 255 is considered to be enzymatically essential, as well as for the causes of the higher reaction rate, the broader substrate specificity, and the lower deamidation activity of this enzyme.Transglutaminase (TGase 1 ; protein-glutamine ␥-glutamyltransferase, EC 2.3.2.13) catalyzes an acyl transfer reaction in which the ␥-carboxyamide groups of peptide-bound glutamine residues act as the acyl donors. The most common acyl acceptors of TGase are the ⑀-amino groups of lysine residues within peptides or the primary amino groups of some naturally occurring polyamines (1, 2). When lysine residues in proteins serve as acyl acceptors, intermolecular or intramolecular ⑀-(␥-glutamyl)lysine bonds are formed, resulting in the polymerization of proteins.TGases are widely distributed in various organisms, including vertebrates (3-7), invertebrates (8, 9), mollusks (10), plants (11), and microorganisms (12). Among these TGases, the human blood coagulation factor XIII has been most characterized (13)(14)(15)(16)(17)(18). By catalyzing the cross-linking between fibrin molecules, factor XIII forms fibrin clots for hemostasis and heals a wound. The crystal structure of human factor XIII has been determined, revealing that it consists of four domains with a cysteine protease-like active site (19 -22). Many TGases are homologous to human factor XIII and share the common feature of Ca 2ϩ -dependent catalytic activity (3-8). A tissue-type TGase from red sea bream liver (fish-derived TGase (FTG)) is an example of such factor XIII-like TGases and shows 33% sequence homology to human factor XIII (7). The crystal structure of FTG has also been determined (23). The overall and active site structures of FTG are essentially similar to those of human factor XIII.A microbial TGase (MTG) has been isolated from the culture medium of Streptoverticillium sp. S-8112 (24), which has been identified as a variant of Sv. mobaraense. This enzyme is the first TGase obtained from a nonmammalian source. Thus far, few TGases have been identified from microorganisms, particularly from Streptoverticillium species (25). Although the physiological role of MTG is still unknown, this protein is secreted from the cytoplas...