TY - JOUR
T1 - Comparison of protein phosphatase inhibition activities and mouse toxicities of microcystins
AU - Chen, Yi Min
AU - Lee, Tzong Huei
AU - Lee, Shyh Jye
AU - Huang, Hsien Bin
AU - Huang, Rang
AU - Chou, Hong Nong
N1 - Funding Information:
We thank Dr Y.E. Chang for instructions in PP-1 and PP-2A activity assays. This work was supported by National Science Council (NSC92-2311-B-002-103 and NSC92-2323-B-002-012), Taiwan, ROC.
PY - 2006/6/1
Y1 - 2006/6/1
N2 - Eight naturally purified microcystins (MCs), including MC-LR, -FR, -WR, -RR, [d-Asp3]MC-FR, -WR, -RR, and [Dha7]MC-RR were utilized to determine the effects of amino acid substitutions and modifications on MC-induced protein phosphatase inhibition activity and mouse toxicity. Catalytic subunits of protein phosphatase 1 (PP-1) and 2A (PP-2A) were purified and subjected to the inhibition assays, and intraperitoneal injection was used to administer MCs into mice for the toxicity assay. It is found that the replacement of the non-polar amino acid l-leucine at the second position of these heptacyclic peptide toxins by a polar l-arginine reduces their mouse toxicities and inhibitory activities against PP-1 and PP-2A to different extends. Demethylation of methyldehydroalanine (Mdha) at the seventh amino acid of MC-RR exhibits the least mouse toxicity and phosphatase inhibition. The loss of a methyl group on the common methylaspartic acid (MeAsp) at the third position of MC-FR, -WR, and -RR does not alter their toxicity levels, but dominantly reduces their activities in PP-1 inhibition compared to other substitutions or modifications. This suggests that the methyl group on MeAsp is also important for MCs inhibition. However, such a tendency is not observed for PP-2A. By comparing the LD50 values of the mouse toxicity assay and IC50 values of the PP-1 and PP-2A inhibition assay of eight MCs using linear regression, it is evident that the MC-induced toxicity is much more related to the inhibition of PP-2A than PP-1, which suggests that PP-2A inhibition may play a major role in the MC-induced mouse toxicity.
AB - Eight naturally purified microcystins (MCs), including MC-LR, -FR, -WR, -RR, [d-Asp3]MC-FR, -WR, -RR, and [Dha7]MC-RR were utilized to determine the effects of amino acid substitutions and modifications on MC-induced protein phosphatase inhibition activity and mouse toxicity. Catalytic subunits of protein phosphatase 1 (PP-1) and 2A (PP-2A) were purified and subjected to the inhibition assays, and intraperitoneal injection was used to administer MCs into mice for the toxicity assay. It is found that the replacement of the non-polar amino acid l-leucine at the second position of these heptacyclic peptide toxins by a polar l-arginine reduces their mouse toxicities and inhibitory activities against PP-1 and PP-2A to different extends. Demethylation of methyldehydroalanine (Mdha) at the seventh amino acid of MC-RR exhibits the least mouse toxicity and phosphatase inhibition. The loss of a methyl group on the common methylaspartic acid (MeAsp) at the third position of MC-FR, -WR, and -RR does not alter their toxicity levels, but dominantly reduces their activities in PP-1 inhibition compared to other substitutions or modifications. This suggests that the methyl group on MeAsp is also important for MCs inhibition. However, such a tendency is not observed for PP-2A. By comparing the LD50 values of the mouse toxicity assay and IC50 values of the PP-1 and PP-2A inhibition assay of eight MCs using linear regression, it is evident that the MC-induced toxicity is much more related to the inhibition of PP-2A than PP-1, which suggests that PP-2A inhibition may play a major role in the MC-induced mouse toxicity.
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U2 - 10.1016/j.toxicon.2006.01.026
DO - 10.1016/j.toxicon.2006.01.026
M3 - Article
C2 - 16684551
AN - SCOPUS:33646786414
SN - 0041-0101
VL - 47
SP - 742
EP - 746
JO - Toxicon
JF - Toxicon
IS - 7
ER -