Characterization of prolidase activity using capillary electrophoresis with tris(2,2'-bipyridyl)ruthenium(II) electrochemiluminescence detection and application to evaluate collagen degradation in diabetes mellitus.
A new method for prolidase (PLD, EC 3.4.13.9) activity assay was developed based on the determination of proline produced from enzymatic reaction through capillary electrophoresis (CE) with tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)3(2+)] electrochemiluminescence detection (ECL). A detection limit of 12.2 fmol (S/N = 3) for proline, corresponding to 1.22 x 10(-8) units of prolidase catalyzing for 1 min was achieved. PLD activity determined by CE-ECL method was in agreement with that obtained from the classical Chinard's one. CE-ECL showed its powerful resolving ability and selectivity as no sample pretreatment was needed and no interference existed. The clinical utility of this method was successfully demonstrated by its application to assay PLD activity in the serum of diabetic patients in order to evaluate collagen degradation in diabetes mellitus (DM). The results indicated that enhanced collagen degradation occurred in DM.
Yuan
Jipei
J
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, China.
Li
Tao
T
Yin
Xue-Bo
XB
Guo
Li
L
Jiang
Xiuzhen
X
Jin
Wenrui
W
Yang
Xiurong
X
Wang
Erkang
E
D015082:2,2'-Dipyridyl / Q000031:analogs & derivatives* / Q000737:chemistry; D003094:Collagen / Q000032:analysis* / Q000378:metabolism; D056831:Coordination Complexes; D003920:Diabetes Mellitus / Q000097:blood* / Q000201:enzymology / Q000378:metabolism; D004150:Dipeptidases / Q000032:analysis*; D004563:Electrochemistry; D019075:Electrophoresis, Capillary / Q000379:methods; D004789:Enzyme Activation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008163:Luminescent Measurements / Q000379:methods*; D011392:Proline / Q000032:analysis; D012680:Sensitivity and Specificity; D013997:Time Factors; D017975:Ruthenium Compounds
D015082:2,2'-Dipyridyl; D003094:Collagen; D056831:Coordination Complexes; D004150:Dipeptidases; D011392:Proline; C547232:tris(2,2-bipyridine)-ruthenium(II); D017975:Ruthenium Compounds; C022449:proline dipeptidase