Charge microheterogeneity of the β-trace proteins (lipocalin-type prostaglandin D synthase) in the cerebrospinal fluid of patients with neurological disorders analyzed by capillary isoelectrofocusing

Abstract

Charge microheterogeneity of the beta-trace protein (beta-TP = lipocalin-type prostaglandin D synthase) in the cerebrospinal fluid (CSF) of patients with various neurological disorders was analyzed by capillary isoelectric focusing (CIEF). Under the conditions employed, beta-TP in the low-molecular-weight protein fraction of CSF was separated into at least four isoforms with different p/ values. An isoform with the pl value of 4.6-4.8 was usually the most abundant. The total beta-TP level in the CSF was determined by enzyme-linked immunosorbent assay (ELISA) to be elevated in patients recovering from organic damage to the CNS and those with pathological brain atrophy. Changes in the total beta-TP level in the CSF were occasionally accompanied by those in its charge microheterogeneity, as revealed by CIEF. Such quantitative and qualitative changes in beta-TP in human CSF indicated changes in its pathophysiological roles in association with various neurological disorders.

Studies on the Properties and Real Existence of Aqueous Solution Systems that are Assumed to Have Antioxidant Activities by the Action of "Active Hydrogen"

“…The reaction mixtures thus prepared were left at 4°C in the dark, and at 0 hr (when the reaction was started), 17 hr, 41 hr and 65 hr after the preparation, bilirubin (BR) was determined by the diazo method as described previously. 7) (3) Rutin (= quercetin-3-rhamnoglucoside, Kanto Chemicals, Tokyo, Japan) was dissolved at a concentration of 100 µM in solutions (1.0 ml each) that were prepared by adding 100 mM phosphate buffer solution (pH 6.8) (0.25 ml each) to the mixtures (0.75 ml each) of same samples as in (2) and DW at volume ratios of 0 : 3 (the control), 1 : 2, 2 : 1 and 3 : 0, respectively. The enzymatic oxidation of rutin was started by mixing the above-mentioned rutin solutions with an equal volume of an aqueous solution in which 25 units of polyphenol oxidase (PPO) (mushroom tyrosinase, Sigma, St. Louis, MO, U.S.A.) was dissolved.…”

Post-translational modification regulates prostaglandin D2 synthase apoptotic activity: Characterization by site-directed mutagenesis

“…Certain isoforms have been reported to be prominent in the CSF of patients with neurological disorders 2-4 . In addition, a variety of cardiovascular implications have been associated with L-PGDS 1, 5-10 or it's enzymatic product, prostaglandin (PG) D 2 [11][12][13][14] . In fact, we recently observed accelerated cardiovascular disease, impaired glucose tolerance, and insulin resistance in L-PGDS knockout mice 15 .…”

Microscale separation methods for enzyme kinetics assays

“…Generally, the one-step procedure enables faster analysis than two-step CIEF. Hiraoka et al [87] successfully used one-step CIEF for rapid determination of microheterogenities of the β-trace protein. Two-step CIEF was used by Shimura et al [88] for trypsin assay and by Hui et al [89] for characterization of cellobiohydrolase I.…”