Substrate requirements for transglutaminases. Influence of the amino acid residue preceding the amine donor lysine in a native protein.
Thirteen recombinant alpha A-crystallin mutants were constructed that differed in the type of amino acid residue directly preceding the sole amine donor lysine for transglutaminases in this protein. The capacity of these mutants to be cross-linked to amine acceptor substrates by tissue transglutaminase and factor XIII was assessed. Two different biotinylated glutamine-containing oligopeptides were used as amine acceptor probes. It appears that the type of residue preceding the amine donor lysine has a considerable influence on the substrate potential of alpha A-crystallin for transglutaminases. This influence shows qualitatively similar trends for tissue transglutaminase and factor XIII and is irrespective of the amine acceptor probe. In general, glycine or aspartic acid before the amine donor lysine has the strongest adverse effects on substrate reactivity, and proline, histidine, and tryptophan are less favorable. Valine, arginine, and phenylalanine, and to a more variable or somewhat lesser extent also serine, alanine, leucine, tyrosine, and asparagine, have an enhancing effect. This pattern of preference is largely in agreement with that observed for the limited number of characterized amine donor lysines in protein substrates for transglutaminases. It can be concluded that tissue transglutaminase and factor XIII have a rather broad yet clearly differentiated tolerance with respect to the residue preceding the amine donor lysine substrate in native proteins.
Grootjans
J J
JJ
Department of Biochemistry, University of Nijmegen, The Netherlands.
Groenen
P J
PJ
de Jong
W W
WW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D003459:Crystallins / Q000737:chemistry / Q000378:metabolism*; D018076:DNA, Complementary; D004926:Escherichia coli; D005174:Factor XII / Q000378:metabolism*; D008239:Lysine; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009842:Oligopeptides / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D015183:Restriction Mapping; D013379:Substrate Specificity; D011503:Transglutaminases / Q000378:metabolism*
D003459:Crystallins; D018076:DNA, Complementary; D009842:Oligopeptides; D011994:Recombinant Proteins; D005174:Factor XII; D011503:Transglutaminases; D008239:Lysine