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Biomedical subjects

Y B Ke

Publications and source records attributed to Y B Ke.

6 recordsLinked to original sources

[Activation of G protein on the membrane of TCS-sensitive cells].

Trichosanthin (TCS), a Type I Ribosome Inactivating Protein isolated from the root tuber of Trichosanthes Kirilowii M. has various biological activities including abortion induction, anti-tumor and anti-HIV. The mechanism of TCS specifically killing sensitive cells has not been studied clearly. In this study, we initially found that there exists TCS-affinity molecule on Syncytiotrophoblast cells and Jar cells. Furthermore, by [35S]GTP gamma S Binding Assay, we find that TCS can activate G protein on the membrane of TCS-sensitive cells. These results indicate that on the membrane of TCS-sensitive cells exists TCS-specific receptor.

Cell Membrane↗

[Study on Trichosanthin induced apoptosis of leukemia K562 cells].

Trichosanthin (TCS), an eukaryotic ribosome-inactivating protein isolated from the root tuber of Trichosanthes plant, has various biological activities including abortion induction, antitumor, and anti-HIV. In this study, cultured human leukemia K562 cells treated with trichosanthin were examined. Analysis of the cells by single laser flow cytometry showed the sub-G1 peak. DNA extracted from these cells formed a characteristic "ladder" on agarose gel electrophoresis. Under electromicroscope, typical morphological changes of apoptosis were also observed. From all of these findings, we concluded that trichosanthin was able to induce apoptosis in K562 cells.

Antineoplastic Agents, Phytogenic↗

Structure-function relationship of trichosanthin.

Trichosanthin (TCS), a 27 kDa ribosome inactivating protein extracted from the root tuber of Trichosanthes plant, was subjected to limited chymotrypsin digestion and three peptide fragments designated TCS-F1 72-247, TCS-F2 100-247, and TCS-F3 133-247 were generated. The RNA-N-glycosidase and cytotoxic activities of the TCS fragments were compared with that of inact TCS. TCS-F1 and TCS-F2 were biologically active, while TCS-F3 was completely inactive. Dose-dependent studies showed that TCS-F1 and TCS-F2 were less potent in their trophoblast cytotoxicity than intact TCS, however, full biological effect could still be obtained with a higher dosage. Based on the known three-dimensional structure of TCS, we postulate that the putative active site of TCS is located at amino acid residues 110 to 174.

Chymotrypsin↗

Structure/function relationship study of Gln156, Glu160 and Glu189 in the active site of trichosanthin.

Trichosanthin is a protein used medicinally in China for abortifacient purposes. It is also an RNA N-glycosidase which inactivates eukaryotic ribosomes by removing adenine4324 from 28S rRNA. Site-directed mutagenesis was performed to probe the role of Gln156, Glu160 and Glu189 in the active site of trichosanthin. The purified altered proteins were assayed for their potency in inhibiting in vitro protein synthesis. The data indicate Glu160 is involved in the catalytic reaction. Kinetics studies suggest the carboxylate group of Glu160 serves to stabilize the transition-state complex. Similar to ricin A, the variant [E160A]trichosanthin is more potent than [E160D]trichosanthin. This is because Glu189 serves as a back-up of the carboxylate group in case Glu160 is mutated to alanine. However, removal of Glu189 in the presence of Glu160 does not affect the ID50 value drastically. An activity of 1800-fold less than that of the wild-type protein was found when both Glu160 and Glu189 were changed to alanine, indicating that some other residues in the active site are also taken part in the lowering of energy barrier for the catalytic reaction. Although Gln156 is highly conserved in related proteins, its mutation to alanine only slightly decreases the activity, showing that this residue does not participate directly in catalysis.

Amino Acid Sequence↗

[4P- syndrome (Wolf-Hirschhorn syndrome) complicated with delay onset of malignant hyperthermia: a case report].

One 8-month-old female patient, weighted 5 kg, with congenital abnormality (4P- syndrome) underwent elective cheiloplasty for cleft lip and palate. Two hours later, with smooth anesthesia and operation, a life-threatening anesthetic complication of malignant hyperthermia occurred at pediatric intensive care unit. The immediate treatments were initially hyperventilating the patient with 100% O2 and cooling the patient with ice bags. Subsequently, intravenous dantrolene 2.5 mg/kg and symptomatic supportive care were administered successfully to treat the event. Upon reviewing the articles, we found that a congenital chromosome 4P deletion abnormality complicated with a delay onset of malignant hyperthermia has not been described previously.

Chromosome Aberrations↗

Variations of gossypol susceptibility in rat spermatozoa during spermatogenesis.

Sperm collected at two epididymal regions from various rat groups force-fed with gossypol were assayed for their ATP levels and motilities to examine their response to the antifertility effect of gossypol, a yellowish polyphenolic pigment from cottonseed oil. Rats receiving gossypol administration for 2 weeks began to show a proportional decrease in spermatozoal ATP content and motility and this antifertility effect deepened with time. Recovery from the gossypol-induced ATP decrease also developed at 2 weeks after the removal of gossypol administration. This rapid development of both the inhibitory and the recovery effects in a normal 53-day spermatogenic process might have stemmed from a change of susceptibility to gossypol inhibition during spermatogenesis. A model showing a more susceptible middle stage in spermatogenesis is proposed.

Adenosine Triphosphate↗