Peptide array on cellulose support--a screening tool to identify peptides with dipeptidyl-peptidase IV inhibitory activity within the sequence of α-lactalbumin.
The inhibition of the enzyme dipeptidyl-peptidase IV (DPP-IV) is an effective pharmacotherapeutic approach for the management of type 2 diabetes. Recent findings have suggested that dietary proteins, including bovine α-lactalbumin, could be precursors of peptides able to inhibit DPP-IV. However, information on the location of active peptide sequences within the proteins is far from being comprehensive. Moreover, the traditional approach to identify bioactive peptides from foods can be tedious and long. Therefore, the objective of this study was to use peptide arrays to screen α-lactalbumin-derived peptides for their interaction with DPP-IV. Deca-peptides spanning the entire α-lactalbumin sequence, with a frame shift of 1 amino acid between successive sequences, were synthesized on cellulose membranes using "SPOT" technology, and their binding to and inhibition of DPP-IV was studied. Among the 114 α-lactalbumin-derived decamers investigated, the peptides 60WCKDDQNPHS69 (αK(i) = 76 µM), 105LAHKALCSEK114 (K(i) = 217 µM) and 110LCSEKLDQWL119 (K(i) = 217 µM) were among the strongest DPP-IV inhibitors. While the SPOT- and traditionally-synthesized peptides showed consistent trends in DPP-IV inhibitory activity, the cellulose-bound peptides' binding behavior was not correlated to their ability to inhibit the enzyme. This research showed, for the first time, that peptide arrays are useful screening tools to identify DPP-IV inhibitory peptides from dietary proteins.
Faculty of Land and Food Systems, Food, Nutrition and Health Program, the University of British Columbia, 2205 East Mall, Vancouver, BC V6T 1Z4, Canada. isablac@mail.ubc.ca.
Isabelle M E
IM
Lacroix
Faculty of Land and Food Systems, Food, Nutrition and Health Program, the University of British Columbia, 2205 East Mall, Vancouver, BC V6T 1Z4, Canada. Eunice.Li-Chan@ubc.ca.
Eunice C Y
EC
Li-Chan
D000595:Amino Acid Sequence; D002482:Cellulose / Q000737:chemistry; D060326:Chemistry Techniques, Synthetic; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism*; D054873:Dipeptidyl-Peptidase IV Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D055808:Drug Discovery; D057809:HEK293 Cells; D006801:Humans; D007768:Lactalbumin / Q000737:chemistry*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology*; D040081:Protein Array Analysis
D054873:Dipeptidyl-Peptidase IV Inhibitors; D010455:Peptides; D002482:Cellulose; D007768:Lactalbumin; D018819:Dipeptidyl Peptidase 4