Mutagenesis study of the glycosylphosphatidylinositol phospholipase C of Trypanosoma brucei.

Abstract:

:The glycosylphosphatidylinositol phospholipase C (GPI-PLC) from Trypanosoma brucei is particularly effective in hydrolysing the GPI-anchors of some proteins. The enzyme is inhibited by Zn2+ and p-chloromercurylphenylsulphonic acid, both of which can act as sulphydryl reagents, suggesting that a cysteine residue may be important in catalysis. Single cysteine to serine mutants have been produced for all eight cysteines in GPI-PLC; all the mutants were fully active in vitro and were still susceptible to p-chloromercurylphenylsulphonic acid inhibition. In contrast, a single histidine 34 to glutamine mutation totally inactivated GPI-PLC. The histidine was chosen after a sequence alignment with the Bacillus cereus phosphatidylinositol phospholipase C (PI-PLC) suggested a conservation of active site residues, including histidine 34 which is central to the proposed reaction mechanism (Heinz D.W., Ryan M., Bullock T.L., Griffith O.H. EMBO J 1995;14:3855-3863). The results suggest that the GPI-PLC and bacterial PI-PLCs have conserved active sites and that the inhibition of GPI-PLC by sulphydryl reagents can occur through more than one residue.

journal_name

Mol Biochem Parasitol

authors

Carnall N,Webb H,Carrington M

doi

10.1016/s0166-6851(97)00177-1

subject

Has Abstract

pub_date

1997-12-15 00:00:00

pages

423-32

issue

2

eissn

0166-6851

issn

1872-9428

pii

S0166-6851(97)00177-1

journal_volume

90

pub_type

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