Open AccessEnzymatic methylation of the Mycobacterium tuberculosis heparin‐binding haemagglutinin
Author(s) -
Host Hélène,
Drobecq Hervé,
Locht Camille,
Menozzi Franco D.
Publication year - 2007
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2007.00636.x
Subject(s) - methylation , mycobacterium tuberculosis , enzyme , proteolysis , biochemistry , recombinant dna , biology , monoclonal antibody , microbiology and biotechnology , chemistry , antibody , tuberculosis , gene , genetics , medicine , pathology
Heparin‐binding haemagglutinin (HBHA) is an important Mycobacterium tuberculosis virulence factor. It displays a complex methylation pattern in its C‐terminal, functional domain, which protects this domain against proteolysis. Here, it is shown that HBHA methylation is catalysed by mycobacterial enzymes and a radio‐enzymatic and a nonradioactive enzyme assay are described, based on the recognition of methylated HBHA by monoclonal antibodies. MS analysis of in vitro methylated HBHA shows a complex methylation pattern similar to that of naturally methylated HBHA. Using recombinant hybrid molecules as acceptor substrates, it was found that the N‐terminal domain of HBHA is not required for recognition by the HBHA‐methyltransferase(s), although it is required for in vivo methylation.