The Journal of Experimental Medicine
Avanti Polar Lipids, Inc.
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
This Article
Right arrow Full Text
Right arrow Full Text (PDF, 144K)
Right arrow PPT slides of all figures
Right arrow Alert me when this article is cited
Right arrow Citation Map
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JEM
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Mühlradt, P. F.
Right arrow Articles by Jung, G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mühlradt, P. F.
Right arrow Articles by Jung, G.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Facebook   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?
© The Rockefeller University Press, 0022-1007/1997/6/1951/ $5.00
The Journal of Experimental Medicine, Volume 185, Number 11, June 2, 1997 1951-1958

Article

Isolation, Structure Elucidation, and Synthesis of a Macrophage Stimulatory Lipopeptide from Mycoplasma fermentans Acting at Picomolar Concentration

Peter F. Mühlradt*, Michael Kieß*, Holger Meyer*, Roderich Süßmuth{ddagger}, and Günther Jung{ddagger}

From the * Immunobiology and Structure Research Groups, Gesellschaft für Biotechnologische Forschung mbH, D-38124 Braunschweig, Germany; and {ddagger} Institut für Organische Chemie der Universität Tübingen, Auf der Morgenstelle 18, D-72076 Tübingen, Germany

Macrophages are typically stimulated by components of microbial cell walls. Surprisingly, cell wall–less mycoplasmas can also very efficiently stimulate macrophages. We showed recently that mycoplasma-derived lipopeptides constitute the active principle. We have now isolated a clone of Mycoplasma fermentans expressing mainly one macrophage-stimulating lipopeptide. This lipopeptide was detergent-extracted and isolated by reversed-phase high-performance liquid chromotography, using nitric oxide release from C3H/HeJ mouse macrophages as bioassay for detection. In contrast to "conventional" bacterial lipoproteins, this lipopeptide had a free NH2 terminus. Amino acid composition, sequence, and the molecular weight of 2,163.3 are consistent with the following structure: S-(2,3-bisacyloxypropyl)cysteine-GNNDESNISFKEK with one mole C16:0, and a further mole of a mixture of C18:0 and C18:1 fatty acid per lipopeptide molecule. The sequence could not be found in either the protein identification resource nor the Swiss Prot data bank. We named this 2-kD lipopeptide, macrophage-activating lipopeptide-2 (MALP-2). Synthetic dipalmitoyl MALP-2 and mycoplasma-derived MALP-2 were compared with the bioassay. Both lipopeptides showed an identical dose dependency with a half-maximal response at 10–11 M concentration. MALP-2 may be one of the most potent natural macrophage stimulators besides endotoxin.

Address correspondence to Dr. Peter F. Mühlradt, Immunobiology Research Group, GBF, Mascheroderweg1, D-38124 Braunschweig, Germany.

Note added in proof. While this work was submitted, we became aware of the study by R.E. Hall, S. Agarwal, D.P. Kestler, J.A. Cobb, K.M. Goldstein, and N.S. Chang, who reported the cloning of a MALP-2–related protein in the Biochemical Journal (Hall, R.E., et al. 1996. Biochem. J. 319:919–927).

1Abbreviations used in this paper: Fmoc, fluorenylmethoxycarbonyl; MALDI, matrix-associated laser desorption/ionization time of flight; MALDI-MS, MALDI mass spectroscopy; MALP-2, 2-kD macrophage-activating lipopeptide; MSA, macrophage stimulatory activity; NO, nitric oxide.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Facebook Facebook   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:



  Home | Help | Feedback | Subscriptions | Archive | Search
TABLE OF CONTENTS