Enzyme Commission (EC) Number: 3.4.14.5
Primary Accession Number: P27487 (DPP4_HUMAN)
DPPIV, dipeptidyl aminopeptidase IV, CD26, glycoprotein GP110, glycylproline aminopeptidase, T cell triggering molecule Tp103, X-PDAP, THAM, and adenosine deaminase binding protein (ADAbp).
Native DPPIV is a ubiquitous type II transmembrane glycoprotein and a serine protease of the S9 prolyl-oligopeptidase family. In vivo, it is synthesized with a signal peptide which functions as the membrane anchoring domain.1,2 There is an 88% sequence homology between the human and porcine kidney enzymes. 3 Both the human and porcine kidney enzymes exist as homodimers with a subunit molecular weight of approx. 30 kDa. The high mannose 100 kDa DPPIV precursor is processed in the Golgi to yield a 124 kDa heavily N-and O-linked mature glycoprotein.4 It is then sorted to the apical membrane through the concerted action of both N- and O-linked glycans and it’s association with lipid microdomains.5 The porcine enzyme contains 18.3% carbohydrates, of which the glycan composition is 0.9% fucose, 3.4% mannose, 5.1% galactose, 8.2% glucosamine, and 0.7% sialic acid.1,2
pI: 5.26 (porcine)
Extinction Coefficient: E1% = 11.5 (280 nm)7 (porcine)
DPPIV is highly expressed on endothelial cells, epithelial cells and lymphocytes.8,9,10 It is also present in plasma in its soluble form.11
DPPIV is involved in the regulation of several important physiological processes: 12,13,14
Natural DPPIV substrates include: 12,13
Natural DPPIV ligands include:12,16,17,18,19
DPPIV inhibitors have been found to improve glucose tolerance and preserve islet function in mice.20 DPPIV inhibitors were found to block entry of HIV-1 or HIV-2 into T lymphoblastoid and monocytoid cell lines.21
DPPIV has a post-proline dipeptidyl aminopeptidase activity that hydrolyzes N-terminal dipeptides from the unsubstituted N-terminus of peptides with the sequence of X-Pro-Z and X-Ala-Z.
The optimum pH for activity is 7.4-8.7.13,22 At pH 7.0 DPPIV exhibits about 45% of maximal activity and at pH 9.6 it exhibits about 90% of maximal activity. Below pH 5.0 the enzyme is essentially inactive.11,23
We determine the enzymatic activity of DPPIV using the chromogenic substrate Gly-Pro-p-nitroanilde. Reported KM values are 0.66 mM for Gly-Pro-2-naphthylamide24 and 1 mM for Ala-Ala-2-naphthylamide.23
Unit Definition: One unit will produce 1.0 µmole of p-nitroaniline from Gly-L-Pro p-nitroanilide per min in 100 mM Tris-HCl at pH 8.0 at 37 °C.
Stopped Rate Spectrophotometric Determination using a Microplate Reader
CONDITIONS: T = 37 °C, pH = 8.0, A405nm, Inc. = 15 minutes
REAGENTS:
(Immediately before use prepare the following in cold reagent A. Dilute to obtain 0.04 - 0.08 units/mL.)
PROCEDURE:
CALCULATIONS:
Plot a standard curve of A405nm versus nmoles of pNA. Calculate nmoles/minute and hence from this determine mmoles/min/mL of enzyme.
FINAL ASSAY CONCENTRATION:
In a 0.2 mL reaction mix, the final concentrations are 100 mM Tris, 0.5 mM Gly-Pro-pNA, and varying amounts of enzyme.
This procedure is for informational purposes. For a current copy of our quality control procedure, please contact our Technical Service Department.
Gly-Pro 4-methoxy-β-naphthylamide
Gly-Pro 4-methoxy-β-naphthylamide
Gly-Pro p-nitroanilide hydrochloride
Gly-Pro p-nitroanilide p-toluenesulfonate salt
Gly-Pro-7-amido-4-methylcoumarin hydrobromide
diisopropyl fluorophosphate
phenylmethanesulfonyl fluoride
Ile-Pro-Ile (Diprotin A)
KR-62436
Dipeptidyl Peptidase, human
Dipeptidyl Peptidase from porcine kidney
The porcine enzyme (Prod. No. D7052) is supplied as a lyophilized powder and is soluble in 100 mM Tris HCl buffer, pH 8.0 (1 vial/mL), yielding a clear, colorless solution. The human enzyme (Prod. No. D4943) is supplied as a solution in 10 mM Tris-HCl pH 7.6, 200 mM NaCl, 1 mM EDTA, 10 % glycerol.
As supplied, these products should be stable for a minimum of one year when stored properly at –20 °C.
To continue reading please sign in or create an account.
Don't Have An Account?