Selective modification by transglutaminase of a glutamine side chain in the hinge region of the histidine-388→glutamine mutant of yeast phosphoglycerate kinase

Abstract

The transglutaminase-catalysed incorporation of putrescine and monodansylcadaverine into yeast phosphoglycerate kinase has been studied. There is little incorporation of the amines into wild-type enzyme, but nearly stoichiometric incorporation into the histidine-388----glutamine mutant enzyme. C.d. studies show that the overall structure of the mutant enzyme is very similar to that of the wild-type enzyme. Incorporation of the amines into the mutant enzyme causes no significant change in its activity. Glutamine-388 was shown, by isolation and sequencing of the modified peptide, to be the site of incorporation of monodansylcadaverine into the mutant enzyme. The specificity of the transglutaminase reaction is discussed in the light of available data.

Addressing substrate glutamine requirements for tissue transglutaminase using substance P analogues

“…When the glutaminyl residue is located in the second or third position away from the N‐terminus, it can be recognized by the enzyme (reviewed in [10]) even when a proline residue precedes the glutamine residue. As reported examples that also follow that feature, we could mention β‐casein [21], analogues S14–S20 in the present work, gliadin peptides [22] or osteonectin where Gln‐3 in the sequence APQQEAL‐ was identified as a major substrate in the TG‐catalyzed cross‐linking of differentiating cartilage [23]. In contrast, glutamine residues having a proline residue in the +1 position [20,24,25] are rarely major targets in the TG‐catalyzed cross‐linking of polypeptide chains.…”

The amphipathic peptide mellitin as a tool to study the membrane-dependent activation of tissue transglutaminase

“…2A). These results are in agreement with the accessibility criterion that identified reactive glutamines on solvent-exposed or flexible areas of the molecule [29,31 ]. Gln 25 is the target residue in the absence or presence of phospholipids which also satisfy tTG substrate requirements that have been recently addressed for small peptides containing adjacent or C-terminal glutamines [32].…”

“…A number of reactive glutamine and/or lysine residues have been identified in peptides and proteins functioning as TGase substrates, but it was not possible to derive a consensus sequence around these residues. Several lines of evidence suggest that the chemical nature of the side chains adjacent to the target aminoacid might not be the sole determinant of specificity and that conformational and other, still poor understood, factors might also play an important role [28][29][30]. In particular, recent studies with TGase 1 suggest that membrane components could affect TGase reaction and residue specificity [13,25].…”

Perturbations in protein dynamics associated with ligand binding to the blood coagulation enzymes thrombin and Factor XIII.

“…Pinpointing FXIIIa substrate specificity remains an elusive goal. Investigations in the past have focused on primary sequence specificity (135)(136)(137) and substrate access to the active site for the Q-containing substrate (138). As far as the K containing substrate, some work has examined sequence specificity surrounding the reactive lysine residue (139,140).…”

“…An ideal substrate for FXIIIa, or any TGase, would probably be located within a pliable region of a protein, such as surface loops or sequences near the N-or Cterminus that can reach into the deeply buried catalytic core (138,144).…”