PMID:8105889

Citation	Phillips, MA, Qin, Q, Mehrpouyan, M and Rice, RH (1993) Keratinocyte transglutaminase membrane anchorage: analysis of site-directed mutants. Biochemistry 32:11057-63
Abstract	Keratinocyte transglutaminase is anchored on the cytosolic side of the plasma membrane by fatty acid thioesterification near the amino terminus, a process which is seen to occur within 30 min of synthesis. The importance of a cluster of five cysteines (residues 47, 48, 50, 51, and 53) where acylation was presumed to occur is now demonstrated by site-directed mutagenesis. Transglutaminase mutants in which the cluster is deleted or the cysteines are all converted to alanine or serine are cytosolic. Partial replacement of the cluster, leaving two contiguous cysteines, is sufficient to confer membrane anchorage, while a single cysteine is only partially effective. As demonstrated with a soluble transglutaminase mutant, membrane anchorage confers susceptibility of the amino-terminal region to phorbol ester-stimulated phosphorylation. Attachment of 105 residues from the transglutaminase amino terminus to involucrin, a highly soluble protein, results in membrane anchorage of the hybrid protein. Attachment of the cysteine cluster alone does not result in membrane attachment of involucrin, but a 32-residue segment containing this cluster is sufficient. Stable transfectants of the human transglutaminase in mouse 3T3 cells are membrane-bound, indicating the fatty acid transacylation is not keratinocyte-specific.
Links	PubMed
Keywords	3T3 Cells/enzymology; Acylation; Amino Acid Sequence; Animals; Base Sequence; Cell Membrane/enzymology; Cysteine/chemistry; Cysteine/genetics; Esterification; Fatty Acids/metabolism; Humans; Keratinocytes/enzymology; Mice; Molecular Sequence Data; Mutagenesis, Site-Directed; Phosphorylation; Tetradecanoylphorbol Acetate/pharmacology; Transfection; Transglutaminases/chemistry; Transglutaminases/genetics; Transglutaminases/metabolism; Tumor Cells, Cultured
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
HUMAN:TGM2		GO:0031226: intrinsic to plasma membrane	ECO:0000315:		C		TGK becomes quickly membrane bound. When mutated, TGK mutant is not membrane bound. Figure 2 identifies the localization of native and mutant using fractionation and then immunoblotting of the immunoprecipitation.	complete CACAO 8332
HUMAN:TGM2	part_of	GO:0031226: intrinsic component of plasma membrane	ECO:0000315: mutant phenotype evidence used in manual assertion		C		Seeded From UniProt	complete

References

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