Addressing substrate glutamine requirements for tissue transglutaminase using substance P analogues.
We have investigated the effect on the substrate requirements for guinea pig liver (tissue) transglutaminase of a set of 11 synthetic glutamine substitution analogues making up the full sequence of the naturally occurring tissue transglutaminase substrate substance P. While a number of peptide sequences derived from proteins that are well-recognized as tissue transglutaminase substrates have been studied, the enzyme activity using substitution analogues of full-length natural substrates has not been investigated as thoroughly. Thus, our set of substance P analogues only differs from one to other by one amino acid mutation while the length (of the peptide) is maintained as in the natural parent peptide. Our results indicate that a glutamine residue is not recognized as substrate by the enzyme whether it is placed at the N- or C-terminal or between two positively charged residues or between two proline residues. To further address the effect on enzyme activity of charged amino acids in the vicinity of the reactive glutamine residue, a new set of synthetic charge replacement analogues of substance P has been also studied. Together, the results have identified new minimal requirements for modification of a particular glutamine residue in a polypeptide chain. It would be of interest to set up a full set of such requirements in order to highlight potential glutamine residues as enzyme targets in the growing list of proteins that are being described as transglutaminase substrates.
Pastor
M T
MT
Department de Bioquímica i Biologia Molecular, Universitat de València, Burjassot, Spain.
Diez
A
A
Pérez-Payá
E
E
Abad
C
C
D000595:Amino Acid Sequence; D000818:Animals; D005973:Glutamine / Q000378:metabolism*; D006168:Guinea Pigs; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D009154:Mutation; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D013373:Substance P / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D011503:Transglutaminases / Q000378:metabolism*
D010446:Peptide Fragments; D005973:Glutamine; D013373:Substance P; D011503:Transglutaminases