STRUCTURAL DEFINITION OF THE NONREDUCING TERMINI OF MANNOSE-CAPPED LAM FROM MYCOBACTERIUM-TUBERCULOSIS THROUGH SELECTIVE ENZYMATIC DEGRADATION AND FAST-ATOM-BOMBARDMENT MASS-SPECTROMETRY
D. Chatterjee et al., STRUCTURAL DEFINITION OF THE NONREDUCING TERMINI OF MANNOSE-CAPPED LAM FROM MYCOBACTERIUM-TUBERCULOSIS THROUGH SELECTIVE ENZYMATIC DEGRADATION AND FAST-ATOM-BOMBARDMENT MASS-SPECTROMETRY, Glycobiology, 3(5), 1993, pp. 497-506
The application of extracellular arabinases from a Cellulomonas sp. an
d fast atom bombardment-mass spectrometry (FAB-MS) provided new insigh
t into the structure of lipoarabinomannan (LAM) of Mycobacterium tuber
culosis, a key molecule in the pathogenesis and physiology of the tube
rcle bacillus. Previously, the non-reducing arabinan ends of LAM from
the virulent (Erdman) strain of M.tuberculosis were shown to be 'cappe
d' by short (alpha1-->2)-linked mannopyranose (Manp)-containing oligos
accharides, a product called ManLAM. The structural relationship betwe
en these Manp units and the underlying arabinofuranose (Araf)-containi
ng arabinan was examined by digesting ManLAM from M.tuberculosis Erdma
n with the Cellulomonas enzyme, resolving fragments by various means a
nd subjecting the derivatized oligoglycosylalditols to FAB-MS. The seq
uences Manp2Araf4, Manp3Araf4 and Manp1-6Araf6 were recognized as the
major terminal motifs. Upon complete structural definition, all of the
Ara6-containing products were shown to be based on a 3,5-linked branc
hed Araf unit, whereas those containing Ara4 were linear. Minor non-ma
nnosylated terminal arrangements containing Ara4-6, branched, linear a
nd cyclical, were also recognized. In addition, the mannan 'core' of M
anLAM was isolated from enzyme digests and shown to contain segments o
f the phosphatidylinositol anchor and a 'stub' of the arabinan side-ch
ain in the form of a 'linker' alpha-Araf-(1-5)-Araf unit attached to C
-2, apparently of the penultimate 2,6-linked Manp residue. The structu
ral unravelling of this complex molecule further substantiates the cas
e for structural and biological similarities to the enterobacterial li
popolysaccharides/lipoglycans and other important 'capped' lipooligome
rs such as the lipooligosaccharides of Neisseria species and the lipop
hosphoglycan of Leishmania promastigotes.