Differences in the post‐translational modification of proteins by polyamines between weakly and highly metastatic B16 melanoma cells

Abstract

The identification of (gamma-glutamyl)polyamines in proteolytic digest of proteins from the cytosolic and particulate fractions of B16-F10 and B16-F10Lr6 cell lines, originating from a spontaneous tumor in C57BL/6 mice, indicates that polyamines are incorporated into melanoma cell proteins by transglutaminases (TGases-EC 2.3.2.13). The levels of spermidine-derived protein cross-links were found to be inversely related with the metastatic potential of the 2 melanoma lines. Characterization of TGase activity in the 2 tumor cell lines showed 3 types of enzyme. The soluble cellular TGase activity (TGase C) was higher, and increased more, during the growth of the least metastasizing clone B16-F10Lr6 than in the B16-F10 line, which is the most metastasizing. Consistently, N1,N8-bis(gamma-glutamyl) spermidine, which is responsible for protein cross-link formation, was present in greater amount in B16-F10Lr6 cells. The enhancement by theophylline of soluble-TGase activity and spermidine-dependent protein cross-links of B16-F10 cells reduced, with linear dose dependence, the ability of these cells to penetrate through human fibronectin-coated membrane in an in vitro assay of invasiveness. Our data confirm and extend earlier observations indicating that the propensity of a tumor to metastasize can be indirectly related to intracellular levels of TGase activity, and provide the basis for some speculation concerning the role of polyamines as modifiers of murine melanoma cell proteins in metastasis.

Tissue transglutaminase in mesenchymal tumour cells

“…Thus, the synthesis of tTG cannot be unfavourable for tumour growth and progression as suggested by many previous reports. [16][17][18][21][22][23][24] Our results are in keeping with a recent study by van Groningen et al 2~ These authors investigated the relation between tTG and the metastatic potential of several human melanoma cell lines. They found that the level of tTG is increased, rather than decreased, in highly metastatic melanoma cells as compared to weakly metastatic ones.…”

Antitumor properties of aloe-emodin and induction of transglutaminase 2 activity in B16–F10 melanoma cells

“…In a previous paper, we provided evidence for the formation and intracellular localization of γ-glutamylpolyamine derivatives in two murine melanoma cell lines with different metastatic potentials. Although the results demonstrated the presence of protein-bound polyamines in these cancer cells, pronounced differences were observed in the two cell lines investigated (Beninati et al 1993). The role of the post-translational modification of ECM and basement membrane proteins with polyamines in the metastatic process, catalyzed by an activated TG2, has been extensively investigated in B16-F10 murine melanoma cells (Lentini et al 2008).…”

Ultrastructural immunolocalization of polyamines in HeLa cells subjected to fast-freezing fixation and freeze substitution

“…In addition to the immunochemical evidence for the existence of membrane PA sites, it has also been demonstrated that PA can interact with membrane proteins by covalent bonds mediated by the action of transglutaminases (TG) (Schrode and Folk 1978). TG incorporate the primary amine of PA into the γ-carboxamide groups of glutamine residues in proteins (Beninati et al , 1993Piacentini and Beninati 1988), and in core histones (Ballestar et al 1996).…”