Proteomics Identification of Acyl-acceptor and Acyl-donor Substrates for Transglutaminase in a Human Intestinal Epithelial Cell Line

Stefania Orrù, Ivana Caputo, Alfonsina D’Amato, Margherita Ruoppolo, Carla Esposito

Journal of Biological Chemistry · 2003 · 69 citations · 40 references

DOIFull text

Open access

Concepts

Abstract

Transglutaminase (TG)-catalyzed cross-linking of both intracellular and extracellular proteins is an important biochemical event. However, increased concentrations of cross-linked proteins have been observed in many disorders. Moreover, TG-catalyzed modification of proteins might generate new self-antigens responsible for the autoimmune response, as in celiac disease. The identification of available substrates may offer an understanding of how the TG-catalyzed post-translational modification has an impact on physiology and disease. We used a proteomic approach to identify TG-modified protein targets in human intestinal epithelial cells to determine the extent to which transglutaminase specifically contributes to celiac disease. Two probes used for which the and a which the approach which to and as for of transglutaminase been targets proteins in of and Transglutaminase (TG)-catalyzed cross-linking of both intracellular and extracellular proteins is an important biochemical event. However, increased concentrations of cross-linked proteins have been observed in many disorders. Moreover, TG-catalyzed modification of proteins might generate new self-antigens responsible for the autoimmune response, as in celiac disease. The identification of available substrates may offer an understanding of how the TG-catalyzed post-translational modification has an impact on physiology and disease. We used a proteomic approach to identify TG-modified protein targets in human intestinal epithelial cells to determine the extent to which transglutaminase specifically contributes to celiac disease. Two probes used for which the and a which the approach which to and as for of transglutaminase been targets proteins in of and used celiac protein used celiac protein a of the post-translational modification of is of and of to an may in the of may as an the of have been in to However, the of is have been and in many the and of the and the has been an is in and in the of and Moreover, has been the extracellular has been in extracellular might on in the of proteins to as substrates for the in may determine the of and to of have been to to the of the of The cross-linking of protein has been to a for the of is the in the intestinal is post-translational of proteins may in celiac is an autoimmune in to a and on the of the is the of in and is to as a in the of in which cells However, the to the of in the of as in the of and the of may an to cells for However, in in the intestinal and how post-translational may to is a of the of the which to the and the is of the intestinal and to an increased of in celiac in both and as as in the extracellular Moreover, in proteins as and substrates in intestinal and in on the identification of substrates of in an a to the of the in a human a for the of the in and in probes for which the and a which the proteomic approach used to identify both and may the of of in substrates is to the of intestinal epithelial cells in human and cells the cells in and cells in and in a and in a cells used the and the the cells a of the and cells used for in and to in and used for in probes used for proteins in which the and a which the for in the of in intracellular cells for the the of in the to which the cells to determine the the cells and and in and and The on for and for the used for in protein cells and in and for and the for The for in a and in the of of of in and in on and to to the the in and for in in for and in the the for the in the The as and the to cells and for in the of in in for and in for the in to the in for observed an and and the of protein protein substrates the of the a as and The proteins and a to the proteins specifically a The protein substrates and to a of the protein the on of protein protein protein protein substrates the for to the of The protein the of the in and and in for and in for in the as The and in for The and a new of the to the The for for of the the in and for and a of the to a and a and to The used in in in a of and in in a of both in a and in a The the the the the the of the proteins the a a and a the and a as The a a of a of in in to in on the The in a a to a The as the of the of and on the and of the the used to and and identify and to the protein of the of the used for the and the of the as protein for the the protein used for the as to as and and both as and as of of cells used for in cells both to a extent and to the and the the and the used as as the probes a approach the intracellular of both substrates in cells the proteins which been to to a and an and is a intracellular of the cells the intracellular in a in of proteins in both of cells and The of proteins in the cells the of However, the cross-linking in cells in the determine the in the of in observed in cells an of and the protein in cells in cells and and Moreover, in the of increased the of the the of a in the of protein of the of and proteins of cells and the in the of the as Moreover, cells and of proteins for of cells and of proteins for of cells and and the to the in cells which the the of the intracellular cells used as a The of proteins might as in the and proteins in cells protein in cells in cells as in and to the substrates in cells the in to the of protein a intracellular and Moreover, the protein to and is the substrates to the and proteins as substrates the to the in the the and the the and protein substrates in the proteins the protein an a to the of the proteins in a the protein The proteins The protein the of the in the of proteins and on a the of proteins an in of and and as in of a the as a The protein substrates in in the and The in the in the of the to in to the in and and and a in the and of the the used an in to the and cells both the and the The the to as as proteins the protein The proteins of in and in and of and of proteins cells and and in the of to a to cells used for an in in the of and of proteins and and the and which the of in the of the the in The the the of the proteins protein and in the and and in the The and the in the and the in the the of the specifically the in the and and the protein and and and and the of transglutaminase protein of protein of the proteins is in of the of transglutaminase protein of protein of the proteins is the in of the is protein and as The for protein and in a of in the as a The of the protein to identify protein an to to the protein of in the of the of a of of the to the and the on the to identification of of and of a of and to identify protein of as protein and protein to identification and to the to protein The of the the which to to The and used to the to the identification of the proteins as the a for a The and of the proteins a protein and the and protein substrates of in both and of the of the in in to the of proteins in the and in the is to the protein of as protein and of both intracellular and is for proteins to The post-translational modification of the to as the of the a in However, in post-translational of proteins may generate as in as a a as a in the of and the of may an to have might have a in the of the in as an of as as the of and substrates have been in the of celiac and in The identification of the substrates of in intestinal may offer an understanding of the of the in the of human used the is a of intestinal in the Moreover, is important to cells used to both and biochemical as the in the to in substrates cells both to a extent and to The to and to the of proteins to as and is the However, the is as the the in which the to the substrates to Moreover, the of proteins in and in the proteins in available in proteomic to identify substrates to and of proteins been is to proteins to substrates of as protein and to the proteins targets proteins in of and of proteins in of protein protein of protein protein protein protein protein protein in a new The is proteins in the of the The intestinal a of cells to the of the an important in both and of epithelial The of the is a of and and to as and the protein to to as the proteins have been to cross-linked the of to an Moreover, has been is of the in the of the in to the cross-linking to of of the a of of in the of of and is to as a Two and to to as is as an protein a of protein to a in is for the of the proteins and in intestinal and in the of of the a of the the the post-translational of proteins in to and proteins important for of proteins proteins as the and and the to the protein and The which is to as a to the protein and in in the of and of and to as substrates for is and the of proteins to as and substrates is is to in the of proteins as substrates is The of as and and may of a the intestinal to in the intestinal proteins in is to as a and as has been is in the in a of used to in to the of the has been is a the and the The the the and a protein on is to and the intestinal However, the of of is might the of is the and an of may in the of is proteins in as the in the of is a in in cells identification a a understanding of how in is to in and cross-linking to to the the of in observed the in both and to of is to a of to the of and cross-linked substrates to the to the of in the increased of as and in in to identify substrates in and of the might a of in the of and is responsible for the in the celiac used celiac protein used celiac protein a of the post-translational modification of is of and of to an may in the of may as an the of have been in to However, the of is have been and in many the and of the and the has been an is in and in the of and Moreover, has been the extracellular has been in extracellular might on in the of proteins to as substrates for the in may determine the of and to of have been to to the of the of The cross-linking of protein has been to a for the of is the in the intestinal is post-translational of proteins may in celiac is an autoimmune in to a and on the of the is the of in and is to as a in the of in which cells However, the to the of in the of as in the of and the of may an to cells for However, in in the intestinal and how post-translational may to is a of the of the which to the and the is of the intestinal and to an increased of in celiac in both and as as in the extracellular Moreover, in proteins as and substrates in intestinal and in on the identification of substrates of in an a to the of the in a human a for the of the in and in probes for which the and a which the proteomic approach used to identify both and may the of of in substrates is to the of intestinal epithelial cells in human and cells the cells in and cells in and in a and in a cells used the and the the cells a of the and cells used for in and to in and used for in probes used for proteins in which the and a which the for in the of in intracellular cells for the the of in the to which the cells to determine the the cells and and in and and The on for and for the used for in protein cells and in and for and the for The for in a and in the of of of in and in on and to to the the in and for in in for and in the the for the in the The as and the to cells and for in the of in in for and in for the in to the in for observed an and and the of protein protein substrates the of the a as and The proteins and a to the proteins specifically a The protein substrates and to a of the protein the on of protein protein protein protein substrates the for to the of The protein the of the in and and in for and in for in the as The and in for The and a new of the to the The for for of the the in and for and a of the to a and a and to The used in in in a of and in in a of both in a and in a The the the the the the of the proteins the a a and a the and a as The a a of a of in in to in on the The in a a to a The as the of the of and on the and of the the used to and and identify and to the protein of the of the used for the and the of the as protein for the the protein used for the as to as and and both as and as of human and cells the cells in and cells in and in a and in a cells used the and the the cells a of the and cells used for in and to in and used for in probes used for proteins in which the and a which the for in the of in intracellular cells for the the of in the to which the cells to determine the the cells and and in and and The on for and for the used for in protein cells and in and for and the for The for in a and in the of of of in and in on and to to the the in and for in in for and in the the for the in the The as and the to cells and for in the of in in for and in for the in to the in for observed an and and the of protein protein substrates the of the a as and The proteins and a to the proteins specifically a The protein substrates and to a of the protein the on of protein protein The protein protein substrates the for to the of The protein the of the in and and in for and in for in the as The and in for The and a new of the to the The for for of the the in and for and a of the to a and a and to The used in in in a of and in in a of both in a and in a The the the the the the of the proteins the a a and a the and a as The a a of a of in in to in on the The in a a to a The as the of the of and on the and of the the used to and and identify and to the protein of the of the used for the and the of the as protein for the the protein used for the as to as and and both as and as of of cells used for in cells both to a extent and to the and the the and the used as as the probes a approach the intracellular of both substrates in cells the proteins which been to to a and an and is a intracellular of the cells the intracellular in a in of proteins in both of cells and The of proteins in the cells the of However, the cross-linking in cells in the determine the in the of in observed in cells an of and the protein in cells in cells and and Moreover, in the of increased the of the the of a in the of protein the of the intracellular cells used as a The of proteins might as in the and proteins in cells protein in cells in cells as in and to the substrates in cells the in to the of protein a intracellular and Moreover, the protein to and is the substrates to the and proteins as substrates the to the in the the and the the and protein substrates in the proteins the protein an a to the of the proteins in a the protein The proteins The protein the of the in the of proteins and on a the of proteins an in of and and as in of a the as a The protein substrates in in the and The in the in the of the to in to the in and and and a in the and of the the used an in to the and cells both the and the The the to as as proteins the protein The proteins of in and in and of and of proteins cells and and in the of to a to cells used for an in in the of and of proteins and and the and which the of in the of the the in The the the of the proteins protein and in the and and in the The and the in the and the in the the of the specifically the in the and and the protein and and and and the of transglutaminase protein of protein of the proteins is in of the of transglutaminase protein of protein of the proteins is the in of the is protein and as The for protein and in a of in the as a The of the protein to identify protein an to to the protein of in the of the of a of of the to the and the on the to identification of of and of a of and to identify protein of as protein and protein to identification and to the to protein The of the the which to to The and used to the to the identification of the proteins as the a for a The and of the proteins a protein and the and protein substrates of in both and of the of the in in to the of proteins in the and in the is to the protein of as protein and of cells used for in cells both to a extent and to the and the the and the used as as the probes a approach the intracellular of both substrates in cells the proteins which been to to a and an and is a intracellular of the cells the intracellular in a in of proteins in both of cells and The of proteins in the cells the of However, the cross-linking in cells in the determine the in the of in observed in cells an of and the protein in cells in cells and and Moreover, in the of increased the of the the of a in the of protein the of the intracellular cells used as a The of proteins might as in the and proteins in cells protein in cells in cells as in and to the substrates in cells the in to the of protein a intracellular and Moreover, the protein to and is the substrates to the and proteins as substrates the to the in the the and the the and protein substrates in the proteins the protein an a to the of the proteins in a the protein The proteins The protein the of the in the of proteins and on a the of proteins an in of and and as in of a the as a The protein substrates in in the and The in the in the of the to in to the in and and and a in the and of the the used an in to the and cells both the and the The the to as as proteins the protein The proteins of in the of the the in The the the of the proteins protein and in the and and in the The and the in the and the in the the of the specifically the in the and and the protein and and and and the protein and as The for protein and in a of in the as a The of the protein to identify protein an to to the protein of in the of the of a of of the to the and the on the to identification of of and of a of and to identify protein of as protein and protein to identification and to the to protein The of the the which to to The and used to the to the identification of the proteins as the a for a The and of the proteins a protein and the and protein substrates of in both and of the of the in The in to the of proteins in the and in the is to the protein of as protein and of both intracellular and is for proteins to The post-translational modification of the to as the of the a in However, in post-translational of proteins may generate as in as a a as a in the of and the of may an to have might have a in the of the in as an of as as the of and substrates have been in the of celiac and in The identification of the substrates of in intestinal may offer an understanding of the of the in the of human used the is a of intestinal in the Moreover, is important to cells used to both and biochemical as the in the to in substrates cells both to a extent and to The to and to the of proteins to as and is the However, the is as the the in which the to the substrates to Moreover, the of proteins in and in the proteins in available in proteomic to identify substrates to and of proteins been is to proteins to substrates of as protein and to the proteins targets proteins in of and of proteins in of protein protein of protein protein protein protein protein protein in a new The is proteins in the of the The intestinal a of cells to the of the an important in both and of epithelial The of the is a of and and to as and the protein to to as the proteins have been to cross-linked the of to an Moreover, has been is of the in the of the in to the cross-linking to of of the a of of in the of of and is to as a Two and to to as is as an protein a of protein to a in is for the of the proteins and in intestinal and in the of of the a of the the the post-translational of proteins in to and proteins important for of proteins proteins as the and and the to the protein and The which is to as a to the protein and in in the of and of and to as substrates for is and the of proteins to as and substrates is is to in the of proteins as substrates is The of as and and may of a the intestinal to in the intestinal proteins in is to as a and as has been is in the in a of used to in to the of the has been is a the and the The the the and a protein on is to and the intestinal However, the of of is might the of is the and an of may in the of is proteins in as the in the of is a in in cells identification a a understanding of how in is to in and cross-linking to to the the of in observed the in both and to of is to a of to the of and cross-linked substrates to the to the of in the increased of as and in in to identify substrates in and of the might a of in the of and is responsible for the in the celiac The of both intracellular and is for proteins to The post-translational modification of the to as the of the a in However, in post-translational of proteins may generate as in as a a as a in the of and the of may an to have might have a in the of the in as an of as as the of and substrates have been in the of celiac and in The identification of the substrates of in intestinal may offer an understanding of the of the in the of human used the is a of intestinal in the Moreover, is important to cells used to both and biochemical as the in the to in substrates cells both to a extent and to The to and to the of proteins to as and is the However, the is as the the in which the to the substrates to Moreover, the of proteins in and in the proteins in available in The proteomic to identify substrates to and of proteins been is to proteins to substrates of as protein and to the proteins targets proteins in of and of proteins in The is proteins in the of the The intestinal a of cells to the of the an important in both and of epithelial The of the is a of and and to as and the protein to to as the proteins have been to cross-linked the of to an Moreover, has been is of the in the of the in to the cross-linking to of of the a of of in the of of and is to as a Two and to to as is as an protein a of protein to a in is for the of the proteins and in intestinal and in the of of the a of the the the post-translational of proteins in to and proteins important for The of proteins proteins as the and and the to the protein and The which is to as a to the protein and in in the of and of and to as substrates for is and the of proteins to as and substrates is is to in the of proteins as substrates is The of as and and may of a the intestinal to in the intestinal proteins in is to as a and as has been is in the in a of used to in to the of the has been is a the and the The the the and a protein on is to and the intestinal However, the of of is might the of is the and an of may in the of is proteins in as the in the of is a in The in cells identification a a understanding of how in is to in and cross-linking to to the the of in observed the in both and to of is to a of to the of and cross-linked substrates to the to the of in the increased of as and in in to identify substrates in and of the might a of in the of and is responsible for the in the celiac

References

40