Methyl Branches Research Articles

Localization of Intrachain Modifications in Bacterial Lipids Via Radical-Directed Dissociation.

Intrachain modifications of membrane glycerophospholipids (GPLs) due to formation of the carbon-carbon double bond (C═C), cyclopropane ring, and methyl branching are crucial for bacterial membrane homeostasis. Conventional collision-induced dissociation (CID) of even-electron ions of GPL favors charge-directed fragmentation channels, and thus little structurally informative fragments can be detected for locating intrachain modifications. In this study, we report a radical-directed dissociation (RDD) approach for characterization of the intrachain modifications within phosphoethanolamines (PEs), a major lipid component in bacterial membrane. In this method, a radical precursor that can produce benzyl or pyridine methyl radical upon low-energy CID at high efficiency is conjugated onto the amine group of PEs. The carbon-centered radical ions subsequently initiate RDD along the fatty acyl chain, producing fragment patterns key to the assignment and localization of intrachain modifications including C═C, cyclopropane rings, and methyl branching. Besides intrachain fragmentation, RDD on the glycerol backbone produces fatty acyl loss as radicals, allowing one to identify the fatty acyl chain composition of PE. Moreover, RDD of lyso-PEs produces radical losses for distinguishing the sn-isomers. The above RDD approach has been incorporated onto a liquid chromatography-mass spectrometry workflow and applied for the analysis of lipid extracts from Escherichia coli and Bacillus subtilis.

Selective branch formation in ethylene polymerization to access precise ethylene-propylene copolymers

Polyolefins with branches produced by ethylene alone via chain walking are highly desired in industry. Selective branch formation from uncontrolled chain walking is a long-standing challenge to generate exclusively branched polyolefins, however. Here we report such desirable microstructures in ethylene polymerization by using sterically constrained α-diimine nickel(II)/palladium(II) catalysts at 30 °C–90 °C that fall into industrial conditions. Branched polyethylenes with exclusive branch pattern of methyl branches (99%) and notably selective branch distribution of 1,4-Me2 unit (86%) can be generated. The ultrahigh degree of branching (>200 Me/1000 C) enables the well-defined product to mimic ethylene-propylene copolymers. More interestingly, branch distribution is predictable and computable by using a simple statistical model of p(1-p)n (p: the probability of branch formation). Mechanistic insights into the branch formation including branch pattern and branch distribution by an in-depth density functional theory (DFT) calculation are elucidated.

A Highly Selective and Sensitive Chiral Derivatization Method for High- Performance Liquid Chromatographic Determination of the Stereoisomer Composition of Natural Products With Chiral Branched Alkyl Chains.

In the study of chiral biologically active compounds such as pheromones, the analysis of the stereoisomer composition is essential to gain more insight into their stereochemical diversity, which affects the pheromone communication channels and therefore the diversification of species. This mini-review summarizes the development of fluorescence derivatization reagents for high-performance liquid chromatographic (HPLC) determination of the absolute configuration and stereoisomer composition of natural products with a chiral branched alkyl chain. The diastereomeric separation of anteiso fatty acids bearing a branched methyl group up to the C-26 position was achieved by reversed-phase HPLC under very low column temperature conditions using (1S,2S)-2-(2,3-anthracenedicarboximido)cyclohexanol as a derivatization reagent, enabling fluorescent detection of these compounds at femtomole levels. This method was also applicable to chiral alcohols and amines with chiral branched methyl groups using similar reagents containing a carboxyl group. These reagents were successfully applied to determine the absolute configurations and stereoisomer composition of the chiral alkyl chain of natural compounds including some insect pheromones, miyakosyne A, and plakoside A. The combination of these reagents and two-dimensional HPLC constitutes a very powerful tool for the analysis of the stereoisomers of natural crude samples. Furthermore, the analysis of some natural bioactive substances using this method demonstrated that natural substances are not always optically pure, consisting instead of stereoisomer mixtures exhibiting stronger activity than optically pure enantiomers. These results cast doubts on the concept of biological homochirality and demonstrate that natural pheromones do not always show the highest activity among all stereoisomers.

Escherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acids

BackgroundA broad diversity of natural and non-natural esters have now been made in bacteria, and in other microorganisms, as a result of original metabolic engineering approaches. However, the fact that the properties of these molecules, and therefore their applications, are largely defined by the structural features of the fatty acid and alcohol moieties, has driven a persistent interest in generating novel structures of these chemicals.ResultsIn this research, we engineered Escherichia coli to synthesize de novo esters composed of multi-methyl-branched-chain fatty acids and short branched-chain alcohols (BCA), from glucose and propionate. A coculture engineering strategy was developed to avoid metabolic burden generated by the reconstitution of long heterologous biosynthetic pathways. The cocultures were composed of two independently optimized E. coli strains, one dedicated to efficiently achieve the biosynthesis and release of the BCA, and the other to synthesize the multi methyl-branched fatty acid and the corresponding multi-methyl-branched esters (MBE) as the final products. Response surface methodology, a cost-efficient multivariate statistical technique, was used to empirical model the BCA-derived MBE production landscape of the coculture and to optimize its productivity. Compared with the monoculture strategy, the utilization of the designed coculture improved the BCA-derived MBE production in 45%. Finally, the coculture was scaled up in a high-cell density fed-batch fermentation in a 2 L bioreactor by fine-tuning the inoculation ratio between the two engineered E. coli strains.ConclusionPrevious work revealed that esters containing multiple methyl branches in their molecule present favorable physicochemical properties which are superior to those of linear esters. Here, we have successfully engineered an E. coli strain to broaden the diversity of these molecules by incorporating methyl branches also in the alcohol moiety. The limited production of these esters by a monoculture was considerable improved by a design of a coculture system and its optimization using response surface methodology. The possibility to scale-up this process was confirmed in high-cell density fed-batch fermentations.

Are Bases Alkylated by Branched Alkylating Agents Removed by Nucleotide Excision Repair or Base Excision Repair?

We have shown that comet-positive responses by adducts that are repaired by nucleotide excision repair (NER) and base excision repair (BER) are reduced by α-amanitin (AMN) and enhanced by 2’, 3’-dideoxythymidine (ddT), respectively. Based on this finding, bases alkylated by n-alkyl groups with ≦5 carbons and those with ≧4 carbons were discussed to be removed by BER and NER, respectively. To study whether bases alkylated by branched alkyl groups can be removed by NER or BER, 4 alkyl methanesulfonates that have a branched methyl group with different branching positions [iso-propyl methanesulfonates (iPMS), 1-methyl-pentyl methanesulfonate (1MPeMS), 2-methyl-pentyl methanesulfonate (2MPeMS), and 3-methyl-pentyl methanesulfonate (3MPeMS)] were synthesized. We studied the reduction of comet-positive responses by AMN and their enhancement by ddT for those alkyl methanesulfonates with a branched methyl group

Profiling of branched-chain fatty acids via nitroxide radical-directed dissociation integrated on an LC-MS/MS workflow.

Chain modifications on fatty acyls, such as methyl branching, are important to modulate the biochemical and biophysical properties of lipids. The current lipid analysis workflows which mainly rely on collisional-induced dissociation (CID) to obtain the structural information of lipids often fail in locating the chain modifications. Radical-directed dissociation (RDD) is a new type of tandem mass spectrometry (MS/MS) method capable of producing intrachain cleavages, thus allowing the detailed characterization of lipid structures. In this study, we have developed an RDD method induced by nitroxide radicals (NO˙) for the analysis of branched-chain fatty acids (BCFAs). Fatty acids (FAs) are first amidated by O-benzylhydroxylamine; MS2 CID of the lithium adduct ion of the derivatized FAs uncages the nitroxide radical, which subsequently initiates RDD along the chain. The location of methyl branching can be determined via characteristic 28 Da spacing due to cleavages on either side of the branching point, with enhanced fragmentation observed toward the carbonyl end. This nitroxide-RDD method has been integrated onto reversed-phase liquid chromatography and applied for the profiling of BCFAs from yak milk powder and pooled human plasma samples. Other than the more often encountered iso- and anteiso-BCFAs, we have identified FA n-5 17 : 0 as a minor component from human plasma, which has been rarely reported before.

Ordered hexagonal columnar liquid crystalline self-assembly of mesogen-free sulfonylated side-chain chiral polyethers and their high dielectric property

To achieve large spontaneous polarization for ferroelectric liquid crystalline polymers, we have designed a series of mesogen-free isotactic polyethers with sulfonylated side chains. Different from prior studies using linear alkyl side chains, in this work a methyl branch was implemented in the alkyl side chains of the chiral polyethers. As a result of the continuous twist from the methyl branches and strong dipole-dipole interactions among neighboring sulfonyl groups, a helical chain conformation with tilted side chains was achieved. When the branched side chains were short (e.g., C5) and double chirality existed in both the main chain and side chains, a crystalline structure was obtained. When the branched side chains were long (e.g., ≥ C8) and single chirality existed in the main chain, an ordered hexagonal columnar liquid crystalline phase was induced. Their high-field dielectric property was studied by electric displacement-electric field (D-E) loop tests; however, no ferroelectric behavior was observed up to 200 MV/m. It was considered that the dipole-dipole interaction among neighboring sulfonyl groups in the helical chains was too strong to allow easy ferroelectric flipping.

Comparative genomic analysis of Methanimicrococcus blatticola provides insights into host adaptation in archaea and the evolution of methanogenesis

Other than the Methanobacteriales and Methanomassiliicoccales, the characteristics of archaea that inhabit the animal microbiome are largely unknown. Methanimicrococcus blatticola, a member of the Methanosarcinales, currently reunites two unique features within this order: it is a colonizer of the animal digestive tract and can only reduce methyl compounds with H2 for methanogenesis, a increasingly recognized metabolism in the archaea and whose origin remains debated. To understand the origin of these characteristics, we have carried out a large-scale comparative genomic analysis. We infer the loss of more than a thousand genes in M. blatticola, by far the largest genome reduction across all Methanosarcinales. These include numerous elements for sensing the environment and adapting to more stable gut conditions, as well as a significant remodeling of the cell surface components likely involved in host and gut microbiota interactions. Several of these modifications parallel those previously observed in phylogenetically distant archaea and bacteria from the animal microbiome, suggesting large-scale convergent mechanisms of adaptation to the gut. Strikingly, M. blatticola has lost almost all genes coding for the H4MPT methyl branch of the Wood–Ljungdahl pathway (to the exception of mer), a phenomenon never reported before in any member of Class I or Class II methanogens. The loss of this pathway illustrates one of the evolutionary processes that may have led to the emergence of methyl-reducing hydrogenotrophic methanogens, possibly linked to the colonization of organic-rich environments (including the animal gut) where both methyl compounds and hydrogen are abundant.

Genome-Scale Analysis of Acetobacterium woodii Identifies Translational Regulation of Acetogenesis.

ABSTRACTAcetogens synthesize acetyl-CoA via the CO2-fixing Wood-Ljungdahl pathway. Despite their ecological and biotechnological importance, their translational regulation of carbon and energy metabolisms remains unclear. Here, we report how carbon and energy metabolisms in the model acetogen Acetobacterium woodii are translationally controlled under different growth conditions. Data integration of genome-scale transcriptomic and translatomic analyses revealed that the acetogenesis genes, including those of the Wood-Ljungdahl pathway and energy metabolism, showed changes in translational efficiency under autotrophic growth conditions. In particular, genes encoding the Wood-Ljungdahl pathway are translated at similar levels to achieve efficient acetogenesis activity under autotrophic growth conditions, whereas genes encoding the carbonyl branch present increased translation levels in comparison to those for the methyl branch under heterotrophic growth conditions. The translation efficiency of genes in the pathways is differentially regulated by 5′ untranslated regions and ribosome-binding sequences under different growth conditions. Our findings provide potential strategies to optimize the metabolism of syngas-fermenting acetogenic bacteria for better productivity.IMPORTANCE Acetogens are capable of reducing CO2 to multicarbon compounds (e.g., ethanol or 2,3-butanediol) via the Wood-Ljungdahl pathway. Given that protein synthesis in bacteria is highly energy consuming, acetogens living at the thermodynamic limit of life are inevitably under translation control. Here, we dissect the translational regulation of carbon and energy metabolisms in the model acetogen Acetobacterium woodii under heterotrophic and autotrophic growth conditions. The latter may be experienced when acetogen is used as a cell factory that synthesizes products from CO2 during the gas fermentation process. We found that the methyl and carbonyl branches of the Wood-Ljungdahl pathway are activated at similar translation levels during autotrophic growth. Translation is mainly regulated by the 5′-untranslated-region structure and ribosome-binding-site sequence. This work reveals novel translational regulation for coping with autotrophic growth conditions and provides the systematic data set, including the transcriptome, translatome, and promoter/5′-untranslated-region bioparts.

Study on effects of molecule structure on exhaust emissions from RCCI engine fueled with low alcohol isomers

Due to the existence of reactivity stratification in mixture of Reactivity Controlled Compression Ignition (RCCI) engine, effects of molecule structure of low alcohols on exhaust emissions may be different. Three kinds of molecule structure, hydroxyl group position (OHP), methyl branches (MB) and straight carbon chain (SCC) are constructed through propanol and butanol isomers, and experimental data of exhaust emissions were drawn from RCCI engine under different oxygen mass contents (RO) and start of injections (SOI). Then, changes of exhaust emissions with OHP, MB, SCC, RO and SOI, and real influence factors and influence sequence of them on emissions are analyzed. The results show that: Although a certain emission changes with a certain impact factor, they may not be correlated with each other, which means the determination of real influencing factors and influence sequence is necessary. This determination is helpful to find out the strongest correlation influencing factors to optimize exhaust emissions. It is worth noting that, different emissions have different real influencing factors. NOX emission and particle mass concentration (PMC) are unrelated to molecule structure. On the contrary, they are only correlated to SOI. Moreover, all emissions are unrelated to OHP, and only HC emission is correlated to MB, while SCC is correlated to multiple emissions, and emission types correlated to Ro and SOI are the most.

Biodegradable thermoplastic copolyester elastomers: Methyl branched PBAmT

Abstract A series of novel biodegradable copolyesters named poly(butylene 3-methyl adipate co-terephthalate) (PBAmT) were synthesized from the monomers of 3-methyl adipic acid (AAm), 1,4-butanediol (BDO), and terephthalic acid (TPA) through a process of esterification and polycondensation. 1H NMR analysis shows that they are random copolymers whose composition can be well controlled by the feed ratio of monomers. From the results of DSC and XRD, the introduction of methyl group successfully destroys the crystallizability of the PBAm chains, thus making it become a relative soft segment compared to PBA, while these random PBAmT copolymers constructed by soft segment PBAm and rigid segment PBT change from semi-crystalline polymers to nearly amorphous polymers as the feed ratio of Am increases. Especially, mechanical tests reveal that the copolymers show outstanding elasticity and rebound resilience with excellent strength. These thermoplastic copolyester elastomers with good performance by simply introduction of branched methyl group on polybutylene adipate terephthalate (PBAT) copolymer chains may well explore the potential application of biodegradable PBAT-based material.

Brockarchaeota, a novel archaeal phylum with unique and versatile carbon cycling pathways

Geothermal environments, such as hot springs and hydrothermal vents, are hotspots for carbon cycling and contain many poorly described microbial taxa. Here, we reconstructed 15 archaeal metagenome-assembled genomes (MAGs) from terrestrial hot spring sediments in China and deep-sea hydrothermal vent sediments in Guaymas Basin, Gulf of California. Phylogenetic analyses of these MAGs indicate that they form a distinct group within the TACK superphylum, and thus we propose their classification as a new phylum, ‘Brockarchaeota’, named after Thomas Brock for his seminal research in hot springs. Based on the MAG sequence information, we infer that some Brockarchaeota are uniquely capable of mediating non-methanogenic anaerobic methylotrophy, via the tetrahydrofolate methyl branch of the Wood-Ljungdahl pathway and reductive glycine pathway. The hydrothermal vent genotypes appear to be obligate fermenters of plant-derived polysaccharides that rely mostly on substrate-level phosphorylation, as they seem to lack most respiratory complexes. In contrast, hot spring lineages have alternate pathways to increase their ATP yield, including anaerobic methylotrophy of methanol and trimethylamine, and potentially use geothermally derived mercury, arsenic, or hydrogen. Their broad distribution and their apparent anaerobic metabolic versatility indicate that Brockarchaeota may occupy previously overlooked roles in anaerobic carbon cycling.

Biophysical Insights into the Spatial Organization and Dynamics of Structurally Complex Mycobacterial Membrane Lipids

The remarkably complex cell wall of mycobacterium tuberculosis (Mtb) forms a formidable barrier to many host stresses and therapeutics. It houses distinct lipids with long hydrocarbon chains, and chemical groups such as trehalose, methyl branches and cyclopropane rings; not found in eukaryotic membranes. The current holistic picture of the Mtb cell envelope from cryo-TEM comprises of peptidoglycan (PG) covalently modified with arabinogalactan polymers that are in turn elaborated with mycolic acids (C60-C90). These mycolic acids form the inner leaflet of the mycomembrane (MM); MM also includes an outer leaflet constituted by a variety of non-covalently associated lipids such as phthiocerol dimycocerosate (PDIM), dimycolyltrehalose (TDM), sulfoglycolipids (SL), phosphatidylinositol mannosides (PIMs), lipoarabinomannan (man-LAM) and DAGs along with phospholipids. The vital role of each Mtb membrane component in forming a spatially complex barrier, and how mycobacteria uses its membrane platforms to impact infection biology remain understudied. in our current work we have characterized the outer membrane (OM) and inner membrane (IM) from Mycobacterium smegmatis by isolating and reconstituting lipids from these regions to form model membranes mimicking the natural cell envelope. Using methods such as atomic force microscopy and spectroscopy, infrared spectroscopy, time-resolved fluorescence spectroscopy, two-photon and confocal microscopy, we provide crucial insights into the lateral organization, fluidity and domain architecture of mycobacterial membrane lipids and its correlation to lipid structure. Our findings may facilitate identification of compounds with membrane perturbing activity against specific membranous regions of mycobacterial envelope. We also foresee our characterized mycobacterial model membrane system to serve as molecularly complete membrane scaffolds for anti-tubercular drug membrane screening in future.

Measurements and simulations of ignition delay times and laminar flame speeds of nonane isomers

Ignition delay times (IDTs) and laminar flame speeds ( S L ) of C 9 H 20 (nonane) isomers are systematically investigated. IDTs of normal nonane (n-C 9 ), 2-methyloctane (2mC 8 ), 2,4-dimethylheptane (24mC 7 ), and 2,2,4,4-tetramethylpentane (2244mC 5 ) are experimetally obtained by a shock tube facility and numerically simulated by a chemkin 0-D reactor model. Further, laminar flame speeds ( S L ) of n-C 9 and 2244mC 5 are measured by spherical expanding flames in a constant-temperature, constant-pressure dual-chamber cruciform burner over a wide range of the equivalence ratio (Φ = 0.7–1.4), which are used to compare with numerically simulated results obtained by chemkin 1-D flame speed model. Detailed reaction mechanisms of KUCRS, LLNL and JetSurF ver.02 are used for numerical simulations. It is found that experimental IDTs increase with the number of methyl branches, especially in low-temperature and negative temperature coefficient (NTC) regions, where the increase of IDT with the number of methyl branches are well predicted by KUCRS. We also find that the measured values of S L of highly branched 2244mC 5 are smaller than those of n-C 9 at all values of Φ studied, of which measured S L data are successfully reproduced by the 1-D flame speed model with KUCRS. These results are important to our understanding of reaction characteristics for highly branched nonane isomers and for the designing of optimal alternative fuels in internal combustion engines.

Characterization of Fatty Acyl Modifications in Phosphatidylcholines and Lysophosphatidylcholines via Radical-Directed Dissociation.

Phosphatidylcholines (PCs) are the major structural components of the plasma membrane of mammalian cells, while lysophosphatidylcholines (LPCs) are critical intermediates in lipid remodeling. Conventional tandem mass spectrometric (MSn) methods via collision-induced dissociation (CID) are blind to intrachain modifications such as the location of the carbon-carbon double bond (C═C) and methyl branching point. In this study, we demonstrate that almost complete structural information can be inferred from a single MS2 CID spectrum of the bicarbonate anion adducts of PC or LPC ([M + HCO3]-), including the identity of the headgroup, composition of fatty acyl chains, their sn-positions, the location of C═C, and the point of methyl branching in fatty acyls. We have integrated this MS2 CID method onto liquid chromatography for the analysis LPCs in human plasma, revealing the existence of multiple sn-isomers, branched chain isomers, and C═C location isomers of LPC.

Branched Chain Lipid Metabolism As a Determinant of the N-Acyl Variation of Streptomyces Natural Products.

Branched-chain fatty acids (BCFA) are encountered in Gram-positive bacteria, but less so in other organisms. The bacterial BCFA in membranes are typically saturated, with both odd- and even-numbered carbon chain lengths, and with methyl branches at either the ω-1 (iso) or ω-2 (anteiso) positions. The acylation with BCFA also contributes to the structural diversity of microbial natural products and potentially modulates biological activity. For the tunicamycin (TUN) family of natural products, the toxicity toward eukaryotes is highly dependent upon N-acylation with trans-2,3-unsaturated BCFA. The loss of the 2,3-unsaturation gives modified TUN with reduced eukaryotic toxicity but crucially with retention of the synergistic enhancement of the β-lactam group of antibiotics. Here, we infer from genomics, mass spectrometry, and deuterium labeling that the trans-2,3-unsaturated TUN variants and the saturated cellular lipids found in TUN-producing Streptomyces are derived from the same pool of BCFA metabolites. Moreover, non-natural primers of BCFA metabolism are selectively incorporated into the cellular lipids of TUN-producing Streptomyces and concomitantly produce structurally novel neo-branched TUN N-acyl variants.

Formation of branched polyethylenes by ethylene homopolymerization using LNiBr2 homo‐ and heterogeneous precatalysts: Interpretation of the polymer structures in comparison with commercial LLDPE

AbstractComparative data on the micro‐structures and properties of branched polyethylenes (BPE) produced via ethylene homopolymerization over homogeneous N,N‐α‐diimine LNiBr2 complexes with different ligand composition (AlEt2Cl as a cocatalyst) and corresponding supported catalysts LNiBr2/SiO2(Al) (Al[iso‐Bu]3 as a cocatalyst) are presented. Noticeable differences were observed between micro‐structures of BPEs obtained using homo‐ and heterogeneous LNiBr2 complexes as catalysts. Supported catalysts produce BPEs with the majority of methyl branches (17–18 CH3(1000 C)−1 characterized by different molecular masses (1800–210 kg mol−1) and molecular weight distributions (Mw[Mn]−1 = 5.9 and 2.6). Thermal and mechanical properties of these BPE samples obtained over supported Ni catalysts are similar to those of commercial LLDPE samples prepared with metallocene and Ziegler‐Natta catalysts.

Theoretical studies on the reaction kinetics of methyl 2-furoate with hydroxyl radical

Methyl 2-furoate (FAME2) is a model for the potential renewable biofuel of dimethyl furan-2,5-dicarboxylate, with the development of its new synthesis method. The potential energy surfaces of H-abstractions and OH-additions between FAME2 and hydroxyl radical (OH) were studied using CCSD(T)/CBS//M062X/cc-pVTZ. The subsequent isomerization and decomposition reactions were also determined for the primary radicals produced. The results showed that H-abstraction on the branched methyl group was the dominant channel and that the OH-addition reactions on the furan ring had a significant pressure dependency. The rate coefficients presented here provide important kinetic data to support future improvement of the combustion mechanism of FAME2, and present a sound basis for further research into practical fuels.

Carbon monoxide and hydrogen (syngas) as a C1-building block for selective catalytic methylation.

A catalytic reaction using syngas (CO/H2) as feedstock for the selective β-methylation of alcohols was developed whereby carbon monoxide acts as a C1 source and hydrogen gas as a reducing agent. The overall transformation occurs through an intricate network of metal-catalyzed and base-mediated reactions. The molecular complex [Mn(CO)2Br[HN(C2H4PiPr2)2]] 1 comprising earth-abundant manganese acts as the metal component in the catalytic system enabling the generation of formaldehyde from syngas in a synthetically useful reaction. This new syngas conversion opens pathways to install methyl branches at sp3 carbon centers utilizing renewable feedstocks and energy for the synthesis of biologically active compounds, fine chemicals, and advanced biofuels.

Resolving Modifications on Sphingoid Base and N-Acyl Chain of Sphingomyelin Lipids in Complex Lipid Extracts.

Sphingomyelins (SMs) are an essential class of lipids widely existing in different organisms. The sphingoid base and N-acyl chain are two building blocks which can undergo different types of modifications during lipogenesis, including desaturation, hydroxylation, and methyl branching. Current lipidomic analysis methods cannot provide detailed information on these structural motifs. Herein, we developed a tandem mass spectrometric method by harnessing radical-directed dissociation (RDD) from collision-induced dissociation (CID) of the bicarbonate anion adduct of SM ([M + HCO3]-). A major RDD channel produced a high-abundance fragment carrying the intact N-acyl chain, termed as "N-acyl fragment", allowing the assignment of the sphingoid base/N-acyl composition and relative quantitation of compositional isomers of SM at high sensitivity. RDD also produced intrachain fragments in lower abundances, which helped localization of methyl branching and hydroxylation in SM. The acetone Paternò-Büchi (PB) reaction was found to be capable of derivatizing the Δ4 carbon-carbon double bond (C═C) in sphingosine (SPH) base and producing C═C diagnostic ions upon CID, albeit at much lower efficiencies than those of the isolated C═C in alkyl chains. A liquid chromatography-mass spectrometry workflow was developed by incorporating MS2 CID of SM via [M + HCO3]- and PB-MS2 CID. The capability of profiling SM with detailed structural information was demonstrated by analyzing complex lipid extracts from porcine brain and Caenorhabditis elegans. These results provided visualization of the sphingoid base/N-acyl compositional isomers of SM lipids and revealed large structural diversity from each sample. These included identification of the sphingadiene base [d18:1(Δ4,14)], C═C location isomers in N-acyls, C-2 hydroxylation of N-acyls, and iso-methyl branched SPH base.