Enhanced activity of variant phospholipase C-delta1 protein (R257H) detected in patients with coronary artery spasm.
BACKGROUND
We recently demonstrated that phospholipase C (PLC)-delta1 activity in cultured skin fibroblasts obtained from patients with coronary spastic angina (CSA) is enhanced. We tested the hypothesis that structural abnormality in PLC-delta1 isoform is a cause of the enhanced activity.


RESULTS
Sequence analysis of the cDNA coding for PLC-delta1 obtained from fibroblasts revealed that one conversion of guanine to adenine (A) was present at nucleotide position 864 in one CSA patient, resulting in the amino acid replacement of arginine 257 by histidine (R257H). The incidence of 864A/A in genomic DNA, analyzed by single-strand conformation polymorphism, was greater in patients with CSA than in male control subjects (6 of 57 patients with CSA versus 1 of 62 control subjects, P<0.05). The activity of the variant PLC-delta1 protein under free calcium concentration between 10(-8) and 10(-7) mol/L was 2-fold higher than that of the wild-type protein. Baseline intracellular calcium concentration ([Ca2+]i) in human embryonic kidney 293 cells transfected with the variant PLC-delta1 was higher than that in cells with the wild type. The peak increase in [Ca2+]i in response to acetylcholine at 10(-6) and 10(-5) mol/L was greater in the cells with the variant PLC-delta1 than in those with the wild type.


CONCLUSIONS
These findings indicate that the R257H variant in the PLC-delta1 gene detected in patients with CSA is associated with enhancement of enzyme activity, and they describe a novel mechanism for the enhanced coronary vasomotility in CSA.
Nakano
Takao
T
Second Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan.
Osanai
Tomohiro
T
Tomita
Hirofumi
H
Sekimata
Masayuki
M
Homma
Yoshimi
Y
Okumura
Ken
K
D000109:Acetylcholine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D003329:Coronary Vasospasm / Q000201:enzymology* / Q000235:genetics; D004247:DNA / Q000032:analysis; D004789:Enzyme Activation; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism; D054803:Phospholipase C delta; D020641:Polymorphism, Single Nucleotide; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D010738:Type C Phospholipases / Q000235:genetics* / Q000378:metabolism*
D007527:Isoenzymes; D019269:Phosphatidylinositol 4,5-Diphosphate; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004247:DNA; D010738:Type C Phospholipases; C518024:PLCD1 protein, human; D054803:Phospholipase C delta; D000109:Acetylcholine; D002118:Calcium