Covalent binding of lipid to protein. Diglyceride and amide-linked fatty acid at the N-terminal end of the murein-lipoprotein of the Escherichia coli outer membrane.

A new structure of a lipid and its covalent linkage to a protein (murein-lipoprotein of the Escherichia coli outer membrane) is described. Glycerylcysteine (S-(propane-2′,3′-diol)-3-thio-2-aminopropanic acid) at the N-terminal end of the polypeptide chain is the attachment site of two ester-bound fatty acids. An additional fatty acid is bound as amide to the N-terminal group. The diglyceride residue on the cysteine could be derived from the phospholipid pathway since the fatty acid composition is very similar to that of the phospholipids from the same cells. In contrast, 65% of the amide-linked fatty acid is palmitate. The main fatty acids are palmitic acid (53%), cis-vaccenic acid (20.7%), 9,10-methylene-hexadecanoic acid (10.6%) and palmitoleic acid (9.4%). The structure of glycerylcysteine was established by chemical degradation of the compound containing the fatty acids, by incorporation studies of [35S]sulfate, [35S]cysteine, [14C]cystine, [methyl-3H]methionine and [2-3H]glycerol and by chemical synthesis. Glycerylcysteine was isolated as a constituent of the peptide (Ser)-Ser-Am-Ala-Lys. The C-terminal part of the lipopeptide sequence overlaps with the N-terminal end of the known sequence of the polypeptide chain. Its N-terminal position was confirmed by isolation of the N-terminal polypeptide fragment (position 1–31) after cleavage with cyanogen bromide. This contained the lipid.

[1]  A. Scanu,et al.  Structural and functional aspects of lipoproteins in living systems. , 1969 .

[2]  J. Rabinowitz,et al.  A procedure for the microdetermination of formic acid in periodate oxidation mixtures. , 1962, Analytical biochemistry.

[3]  L. D. Metcalfe,et al.  Rapid Preparation of Fatty Acid Esters from Lipids for Gas Chromatographic Analysis. , 1966 .

[4]  E. P. Kennedy,et al.  Pathways for the synthesis of glycerophosphatides in Escherichia coli. , 1967, The Journal of biological chemistry.

[5]  K. Narita,et al.  On the Hydrazinolysis of Proteins and Peptides: A Method for the Characterization of Carboxyl-terminal Amino Acids in Proteins , 1952 .

[6]  J. Folch-Pi,et al.  On the type of linkage binding fatty acids present in brain white matter proteolipid apoprotein. , 1971, Biochemical and biophysical research communications.

[7]  R. D. Mackenzie,et al.  Rapid colorimetric micromethod for free fatty acids. , 1967, Journal of lipid research.

[8]  V. Braun,et al.  Supramolecular structure of the rigid layer of the cell wall of Salmonella, Serratia, Proteus, and Pseudomonas fluorescens. Number of lipoprotein molecules in a membrane layer. , 1970, Biochemistry.

[9]  E. Rietschel,et al.  Nature and linkages of the fatty acids present in the lipid-A component of Salmonella lipopolysaccharides. , 1972, European journal of biochemistry.

[10]  V. Braun,et al.  Evidence for a Covalent Bond between Lipid and a Membrane Protein (Rigid Layer of the Cell Wall of E. coli) , 1972 .

[11]  D. A. Smith.  S-amino acid metabolism and its regulation in Escherichia coli and Salmonella typhimurium. , 1971, Advances in genetics.

[12]  T. E. Thompson,et al.  Artifacts produced by boron trifluoride methanolysis of a synthetic lecithin containing cyclopropane fatty acids (1-2-dihydrosterculoyl-3-sn-phosphatidylcholine). , 1971, Journal of lipid research.

[13]  W. Fisher,et al.  Structure of Lipoproteins: Covalently Bound Fatty Acids , 1964, Science.

[14]  R. A. Klein.  Mass spectrometry of the phosphatidylcholines: dipalmitoyl, dioleoyl, and stearoyl-oleoyl glycerylphosphorylcholines. , 1971, Journal of lipid research.

[15]  V Braun,et al.  Repetitive sequences in the murein-lipoprotein of the cell wall of Escherichia coli. , 1972, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. Cronan,et al.  Metabolism and function of the membrane phospholipids of Escherichia coli. , 1972, Biochimica et biophysica acta.

[17]  B. Witkop,et al.  Nonenzymatic cleavage of peptide bonds: the methionine residues in bovine pancreatic ribonuclease. , 1962, The Journal of biological chemistry.

[18]  B. D. Davis.  Diaminopimelic Acid and Lysine: Biosynthetic Interrelations of Lysine, Diaminopimelic Acid, and Threonine in Mutants of Escherichia coli , 1952, Nature.

[19]  E. H. Ahrens,et al.  The separation of complex lipide mixtures by the use of silicic acid chromatography. , 1958, The Journal of biological chemistry.

[20]  Gesellschaft für Biologische Chemie. Colloquium,et al.  Inhibitors Tools in Cell Research , 1969, Colloquium der Gesellschaft für Biologische Chemie in Mosbach Baden.

[21]  V. Braun,et al.  Sequence of the murein-lipoprotein and the attachment site of the lipid. , 1972, European journal of biochemistry.

[22]  D. Tipper,et al.  Structure of the cell wall peptidoglycan of Lactobacillus casei RO94. , 1969, Biochemistry.

[23]  J. Strominger.  Penicillin-sensitive enzymatic reactions in bacterial cell wall synthesis. , 1968, Harvey lectures.

[24]  W. Gray.  [12] Dansyl chloride procedure , 1967 .

[25]  W. Morgan,et al.  The determination of glucosamine and galactosamine. , 1955, The Biochemical journal.

[26]  G. Gerisch,et al.  Degradation of the murein-lipoprotein complex of Escherichia coli cell walls by Dictyostelium amoebae. , 1972, European journal of biochemistry.

[27]  R. A. Klein.  Mass spectrometry of the phosphatidylcholines: fragmentation processes for dioleoyl and stearoyl-oleoyl glycerylphosphorylcholine. , 1971, Journal of lipid research.

[28]  E. Lederer,et al.  Determination of amino acid sequences in oligopeptides by mass spectrometry. I. The structure of fortuitine, an acyl-nonapeptide methyl ester , 1965 .

[29]  G. Braunitzer,et al.  The Hæmoglobin Particles , 1961, Nature.

[30]  H. Rogers.  Bacterial growth and the cell envelope. , 1970, Bacteriological reviews.

[31]  J. Ghuysen,et al.  The peptide N alpha-(L-alanyl-D-isoglutaminyl)-N epsilon-(D-isoasparaginyl)-L-lysyl-D-alanine and the disaccharide N-acetylglucosaminyl-beta-1,4-N-acetylmuramic acid in cell wall peptidoglycan of Streptococcus faecalis strain ATCC 9790. , 1967, Biochemistry.

[32]  V. Braun,et al.  Chemical characterization, spatial distribution and function of a lipoprotein (murein-lipoprotein) of the E. coli cell wall. The specific effect of trypsin on the membrane structure. , 1969, European journal of biochemistry.

[33]  V. Braun,et al.  The Covalent Murein‐Lipoprotin Structure of the Escherichia coli Cell Wall , 1970 .

[34]  H. I. Waterman,et al.  On the Mechanism of the Phenyl Isothiocyanate Degradation of Peptides. , 1956 .

[35]  H. Jackson,et al.  Antifertility Effects and Metabolism of α and epi-Chlorhydrins in the Rat , 1969, Nature.

[36]  V. Braun,et al.  A reinvestigation of the hydrazinolytic procedure for the determination of C-terminal amino acids , 1967 .

[37]  F. Smith,et al.  COLORIMETRIC METHOD FOR DETER-MINATION OF SUGAR AND RELATED SUBSTANCE , 1956 .

[38]  A. D. Brown,et al.  ASPECTS OF BACTERIAL RESPONSE TO THE IONIC ENVIRONMENT. , 1964, Bacteriological reviews.

[39]  L. Leive,et al.  The transport of diaminopimelate and cystine in Escherichia coli. , 1965, The Journal of biological chemistry.

[40]  H. H. Martin.  Biochemistry of bacterial cell walls. , 1966, Annual review of biochemistry.

[41]  V. Braun,et al.  The murein-lipoprotein linkage in the cell wall of Escherichia coli. , 1970, European journal of biochemistry.