Influence of dipeptidyl peptidase IV on enzymatic properties of adenosine deaminase.
The importance of ADA (adenosine deaminase) in the immune system and the role of its interaction with an ADA-binding cell membrane protein dipeptidyl peptidase IV (DPPIV), identical to the activated immune cell antigen, CD26, has attracted the interest of researchers for many years. To investigate the specific properties in the structure-function relationship of the ADA/DPPIV-CD26 complex, its soluble form, identical to large ADA (LADA), was isolated from human blood serum, human pleural fluid and bovine kidney cortex. The kinetic constants (Km and Vmax) of LADA and of small ADA (SADA), purified from bovine lung and spleen, were compared using adenosine (Ado) and 2'-deoxyadenosine (2'-dAdo) as substrates. The Michaelis constant, Km, evidences a higher affinity of both substrates (in particular of more toxic 2'-dAdo) for LADA and proves the modulation of toxic nucleoside neutralization in the extracellular medium due to complex formation between ADA and DPPIV-CD26. The values of Vmax are significantly higher for SADA, but the efficiency, Vmax/Km, in LADA-catalyzed 2'-dAdo deamination is higher than that in Ado deamination. The interaction of all enzyme preparations with derivatives of adenosine and erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was studied. 1-DeazaEHNA and 3-deazaEHNA demonstrate stronger inhibiting activity towards LADA, the DPPIV-CD26-bound form of ADA. The observed differences between the properties of the two ADA isoforms may be considered as a consequence of SADA binding with DPPIV-CD26. Both SADA and LADA indicated a similar pH-profile of adenosine deamination reaction with the optimum at pHs 6.5-7.5, while the pH-profile of dipeptidyl peptidase activity of the ADA/DPPIV-CD26 complex appeared in a more alkaline region.
Sharoyan
Svetlana
S
H.Ch. Buniatyan Institute of Biochemistry, Yerevan, Armenia.
Antonyan
Alvard
A
Mardanyan
Sona
S
Lupidi
Giulio
G
Cristalli
Gloria
G
D000241:Adenosine / Q000494:pharmacology; D000243:Adenosine Deaminase / Q000378:metabolism*; D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D002417:Cattle; D016048:Dideoxyadenosine / Q000494:pharmacology; D018819:Dipeptidyl Peptidase 4 / Q000097:blood* / Q000235:genetics; D006801:Humans; D011485:Protein Binding; D013379:Substrate Specificity; D013601:T-Lymphocytes / Q000201:enzymology*
D000945:Antigens, Differentiation, T-Lymphocyte; D016048:Dideoxyadenosine; D018819:Dipeptidyl Peptidase 4; D000243:Adenosine Deaminase; D000241:Adenosine