TY - JOUR
T1 - Structural definition of trehalose 6-monomycolates and trehalose 6,6'-dimycolates from the pathogen Rhodococcus equi by multiple-stage linear ion-trap mass Spectrometry with electrospray ionization
AU - Hsu, Fong Fu
AU - Wohlmann, Jens
AU - Turk, John
AU - Haas, Albert
PY - 2011/12
Y1 - 2011/12
N2 - The cell wall of the pathogenic bacterium Rhodococcus equi (R. equi) contains abundant trehalose monomycolate (TMM) and trehalose dimycolate (TDM), the glycolipids bearing mycolic acids. Here, we describe multiple-stage (MS n) linear ion-trap (LIT) mass spectrometric approaches toward structural characterization of TMM and TDM desorbed as [M + Alk] + (Alk = Na, Li) and as [M + X] - (X = CH 3CO 2, HCO 2) ions by electrospray ionization (ESI). Upon MS n (n=2, 3, 4) on the [M + Alk] + or the [M + X] - adduct ions of TMM and TDM, abundant structurally informative fragment ions are readily available, permitting fast assignment of the length of the meromycolate chain and of the α-branch on the mycolyl residues. In this way, structures of TMM and TDM isolated from pathogenic R. equi strain 103 can be determined. Our results indicate that the major TMM and TDM molecules possess 6, and/or 6'-mycolyl groups that consist of mainly C14 and C16 α-branches with meromycolate branches ranging from C18 to C28, similar to the structures of the unbound mycolic acids found in the cell envelope. Up to 60 isobaric isomers varying in chain length of the α-branch and of the meromycolate backbone were observed for some of the TDM species in the mixture. This mass spectrometric approach provides a direct method that affords identification of various TMM and TDM isomers in a mixture of which the complexity of this lipid class has not been previously reported using other analytical methods.
AB - The cell wall of the pathogenic bacterium Rhodococcus equi (R. equi) contains abundant trehalose monomycolate (TMM) and trehalose dimycolate (TDM), the glycolipids bearing mycolic acids. Here, we describe multiple-stage (MS n) linear ion-trap (LIT) mass spectrometric approaches toward structural characterization of TMM and TDM desorbed as [M + Alk] + (Alk = Na, Li) and as [M + X] - (X = CH 3CO 2, HCO 2) ions by electrospray ionization (ESI). Upon MS n (n=2, 3, 4) on the [M + Alk] + or the [M + X] - adduct ions of TMM and TDM, abundant structurally informative fragment ions are readily available, permitting fast assignment of the length of the meromycolate chain and of the α-branch on the mycolyl residues. In this way, structures of TMM and TDM isolated from pathogenic R. equi strain 103 can be determined. Our results indicate that the major TMM and TDM molecules possess 6, and/or 6'-mycolyl groups that consist of mainly C14 and C16 α-branches with meromycolate branches ranging from C18 to C28, similar to the structures of the unbound mycolic acids found in the cell envelope. Up to 60 isobaric isomers varying in chain length of the α-branch and of the meromycolate backbone were observed for some of the TDM species in the mixture. This mass spectrometric approach provides a direct method that affords identification of various TMM and TDM isomers in a mixture of which the complexity of this lipid class has not been previously reported using other analytical methods.
KW - Cord factor
KW - ESI
KW - Glycolipids
KW - Internal glucose loss
KW - Mycolic acids
KW - Rhodococcus equi
KW - Tandem mass spectrometry
KW - Trehalose dimycolate
KW - Trehalose monomycolate
UR - http://www.scopus.com/inward/record.url?scp=84856205361&partnerID=8YFLogxK
U2 - 10.1007/s13361-011-0240-7
DO - 10.1007/s13361-011-0240-7
M3 - Article
C2 - 21972013
AN - SCOPUS:84856205361
VL - 22
SP - 2160
EP - 2170
JO - Journal of the American Society for Mass Spectrometry
JF - Journal of the American Society for Mass Spectrometry
SN - 1044-0305
IS - 12
ER -