Identification of S-(2,3-dihydroxypropyl)cystein in a macrophage-activating lipopeptide from Mycoplasma fermentans

被引:43
作者
Muhlradt, PF
Meyer, H
Jansen, R
机构
[1] GBF, Gesell. F. Biotechnologische F., D-38124 Braunschweig
关键词
D O I
10.1021/bi9602831
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Mycoplasmas are capable of stimulating monocytes and macrophages to release cytokines, prostaglandins, and nitric oxide. The aim of this study was to characterize the chemical nature of the previously isolated [Muhlradt, P. F., & Frisch, M. (1994) Infect. Immun. 62, 3801-3807] macrophage-stimulating material ''MDHM'' from Mycoplasma fermentans. Mycoplasmas were delipidated, and MDHM activity was extracted with octyl glucoside and further purified by reversed-phase HPLC. Macrophage- stimulating activity was monitored by nitric oxide release from peritoneal macrophages from C3H/HeJ endotoxin low responder mice. HPLC-purified MDHM was rechromatographed on an analytic scale RP 18 column before and after proteinase K treatment. Proteinase treatment did not diminish biological activity but shifted MDHM elution toward higher lipophilicity, suggesting that the macrophage-stimulating activity might reside in the lipopeptide moiety of a lipoprotein. Proteinase K-treated MDHM was hydrolyzed, amino groups were dansylated, and the dansylated material was isolated by HPLC. Dansylated S-(2,3-dihydroxypropyl)cystein (glycerylcystein thioether), typical for Braun's murein lipoprotein, and Dns-Gly and Dns-Thr were identified by tandem mass spectrometry. These amino acids were isolated from biologically active but not from the neighboring inactive HPLC fractions. IR spectra from proteinase K-treated, HPLC-purified MDHM and those from the synthetic lipopeptide [2,3-bis(palmitoyloxy)-(2-RS)-propyl]-N-palmitoyl-(R)-CysSerSerAsnAla were very similar. The data, taken together, indicate that lipoproteins of a nature previously detected in eubacteria are expressed in M. fermentans and that at least one of these lipoproteins and a lipopeptide derived from it constitute the macrophage-activating principle MDHM from these mycoplasmas.
引用
收藏
页码:7781 / 7786
页数:6
相关论文
共 52 条
[41]  
SEID RC, 1980, INFECT IMMUN, V29, P990
[42]   INVITRO INDUCTION OF TUMOR NECROSIS FACTOR-ALPHA, TUMOR CYTOLYSIS, AND BLAST-TRANSFORMATION BY SPIROPLASMA MEMBRANES [J].
SHER, T ;
YAMIN, A ;
ROTTEM, S ;
GALLILY, R .
JOURNAL OF THE NATIONAL CANCER INSTITUTE, 1990, 82 (13) :1142-1145
[43]   MODULATION OF MYELOPOIESIS BY DIFFERENT BACTERIAL CELL-WALL COMPONENTS - INDUCTION OF COLONY-STIMULATING ACTIVITY (BY PURE PREPARATIONS, LOW-MOLECULAR-WEIGHT DEGRADATION PRODUCTS, AND A SYNTHETIC LOW-MOLECULAR ANALOG OF BACTERIAL CELL-WALL COMPONENTS) INVITRO [J].
STABER, FG ;
GISLER, RH ;
SCHUMANN, G ;
TARCSAY, L ;
SCHLAFLI, E ;
DUKOR, P .
CELLULAR IMMUNOLOGY, 1978, 37 (01) :174-187
[44]   MAMMALIAN NITRATE BIOSYNTHESIS - MOUSE MACROPHAGES PRODUCE NITRITE AND NITRATE IN RESPONSE TO ESCHERICHIA-COLI LIPOPOLYSACCHARIDE [J].
STUEHR, DJ ;
MARLETTA, MA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1985, 82 (22) :7738-7742
[45]   LIPOPROTEINS OF GRAM-POSITIVE BACTERIA [J].
SUTCLIFFE, IC ;
RUSSELL, RRB .
JOURNAL OF BACTERIOLOGY, 1995, 177 (05) :1123-1128
[46]   MACROPHAGE-ACTIVATING FACTOR EXTRACTED FROM MYCOPLASMAS [J].
TAKEMA, M ;
OKA, S ;
UNO, K ;
NAKAMURA, S ;
ARITA, H ;
TAWARA, K ;
INABA, K ;
MURAMATSU, S .
CANCER IMMUNOLOGY IMMUNOTHERAPY, 1991, 33 (01) :39-44
[47]   PROFILING AND QUANTIFICATION OF BIOGENIC-AMINES IN WINE BY CONSTANT B2E LINKED SCAN MASS-SPECTROMETRY [J].
WALTHER, H ;
SCHLUNEGGER, UP ;
FRIEDLI, F .
BIOMEDICAL AND ENVIRONMENTAL MASS SPECTROMETRY, 1987, 14 (05) :229-233
[48]   SEPARATION OF DNS-AMINO ACIDS USING REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY - A SENSITIVE METHOD FOR DETERMINING N-TERMINI OF PEPTIDES AND PROTEINS [J].
WEINER, S ;
TISHBEE, A .
JOURNAL OF CHROMATOGRAPHY, 1981, 213 (03) :501-506
[49]   A FAMILY OF STRAIN-VARIANT SURFACE LIPOPROTEINS OF MYCOPLASMA-FERMENTANS [J].
WISE, KS ;
KIM, MF ;
THEISS, PM ;
LO, SC .
INFECTION AND IMMUNITY, 1993, 61 (08) :3327-3333
[50]   MAJOR MEMBRANE-SURFACE PROTEINS OF MYCOPLASMA-HYOPNEUMONIAE SELECTIVELY MODIFIED BY COVALENTLY BOUND LIPID [J].
WISE, KS ;
KIM, MF .
JOURNAL OF BACTERIOLOGY, 1987, 169 (12) :5546-5555