Isolation and studies of the granules of the amebocytes of Limulus polyphemus, the horseshoe crab

  title={Isolation and studies of the granules of the amebocytes of Limulus polyphemus, the horseshoe crab},
  author={Erik H. M{\"u}rer and Jack Levin and R Holme},
  journal={Journal of Cellular Physiology},
Granules were isolated from the cytoplasm of the amebocytes of Limulus polyphemus, the horseshoe crab, by disruption of cells obtained from blood which had been drawn into 2 mM propranolol. The granules subsequently were purified by centrifugation through a sucrose gradient that contained heparin. Extracts of the granules were prepared by freezing and thawing the granule preparations in distilled water. 
Bactericidal activity of granules isolated from amoebocytes of the horseshoe crab, Limulus polyphemus.
Granules isolated from amoebocytes of Limulus polyphemus used in bactericidal studies exhibited marked killing activity against Escherichia coli and Staphylococcus aureus, supporting the idea that amoEBocytes are an important component in the defense system of this animal.
The fine structure of the amebocyte (hemocyte) in the blood of Limulus polyphemus was reinvestigated regarding its normal state to resolve existing discrepancies in reporting before proceeding to
Characterization of amebocyte coagulogen from the horseshoe crab (Limulus polyhemus).
Findings, plus NH2-terminal sequence analysis indicating a single sequence Gly-Asp-Pro-Asn-Val-Pro-, are consistent with a molecular structure comprised of two identical chains joined by non-covalent forces.
Two types of hemocytes in localization of clottable protein in Japanese horseshoe crab, Tachypleus tridentatus
The two types of hemocytes which showed different intracellular localization of CP might be in different maturational stages or might play different roles in blood coagulation.
Proteinase inhibitory activity released from the horseshoe crab blood cell during exocytosis.
The blood cell of the horseshoe crab, Limulus, is packed with granules that can be stimulated to release their contents by exocytosis. We have identified a family of proteinase inhibitors in the
The role of the haemocytes of Clitumnus extradentatus in haemolymph coagulation
  • A. Rowley
  • Biology, Medicine
    Cell and Tissue Research
  • 2004
SummaryLight and electron-microscopic observations of the blood cells (haemocytes) of the stick insect Clitumnus extradentatus in vitro showed that two morphologically distinct cell types, the
Blood Collection from the American Horseshoe Crab, Limulus Polyphemus
The horseshoe crab has the best-characterized immune system of any long-lived invertebrate and its blood clotting system involves aggregation of amebocytes and the formation of an extracellular clot of a protein, coagulin, which is released from the secretory granules of the blood cells.
Morphology of the granular hemocytes of the Japanese horseshoe crabTachypleus tridentatus and immunocytochemical localization of clotting factors and antimicrobial substances
SummaryThe structure of hemocytes in the normal state and during blood coagulation, and the intracellular localization of three clotting factors and two antimicrobial factors were examined in the
Discovery and Early Development of the Limulus Test
  • J. Levin
  • Medicine
    Endotoxin Detection and Control in Pharma, Limulus, and Mammalian Systems
  • 2019
The discovery by the author that the blood coagulation mechanism of Limulus polyphemus, the horseshoe crab, was exquisitely sensitive to bacterial endotoxins provided the basis for the invention of the Limulus amebocyte lysate (LAL) test for the detection of bacterial endotoxin.
Response of the blood clotting system of the American horseshoe crab, Limulus polyphemus, to a novel form of lipopolysaccharide from a green alga.
It is shown that, like LPS from Gram-negative bacteria, the algal molecule stimulates exocytosis of the Limulus blood cell and the clotting of coagulin, and the coagulogen clot efficiently entraps the cells of Chlorella in a network of fibrils.


Some cytologic characteristics of the hemocytes of Limulus during clotting
The hemolymph of the horseshoe crab, Limulus (Xiphosura) polyphemus, contains a single cell type and phase contrast time‐lapse cinematography reveals that the cells are extremely motile during this phase.
An invertebrate coagulation system activated by endotoxin: evidence for enzymatic mediation.
The data suggest that endotoxin activates an enzyme which then gels the clottable protein contained in amebocyte lysate, suggesting that enzymatic activity depended upon serine hydroxyl and sulfhydryl groups.
The coagulogen of Limulus polyphemus hemocytes. A comparison of the clotted and non-clotted forms of the molecule
Abstract The coagulogen (clottable protein) found in lysates of Limulus polyphemus hemocytes has been studied. Purified coagulogen is clotted by endotoxin in the presence of a high molecular weight
Detection of endotoxin in human blood and demonstration of an inhibitor.
The increase in positive tests, following dilution or chloroform extraction of serum or plasma, suggested that the inhibitory nature of blood is primarily the result of reversible binding between endotoxin and serum protein(s), rather than destruction of endotoxin.
Leukocytic proteases and the immunologic release of lysosomal enzymes.
Lvsosomal proteases present in cartilage and in leukocvtes have been shown capable of degrading cartilage matrix, isolated proteoglvcans and simple chemical substrates.
Extraction and Isolation of Individual Ribosomal Proteins from Escherichia coli
By the methods described here, it is possible to isolate in high purity at least 15 different proteins from the 30S ribosome of Escherichia coli.
Fibrinogen associated with subcellular platelet particles.
The subcellular localization of fibrinogen in platelets washed in an EDTA-buffer medium has been studied and results indicate a preferential association of granule-bound fibr inogen with low-density granules.
Immunological Studies of Proteins Associated with the Subcellular Fractions of Thrombasthenic and Afibrinogenaemic Platelets
The results suggest that decreased platelet fibrinogen in thrombasthenia may be a secondary phenomenon, perhaps related to an underlying membrane defect, and supports the concept of two separate pools of fibrInogen—intracellular and extracellular.
Compounds known to affect the cyclic adenosine monophosphate level in blood platelets: effect on thrombin-induced clot retraction and platelet release.
  • E. Mürer
  • Chemistry, Medicine
    Biochimica et biophysica acta
  • 1971
It is concluded that compounds which are knwon to increase the cyclic AMP level in blood platelets exert an inhibitory effect on clot retraction and the platelet release reaction, and that the inhibition is counteracted when the impulse for platelet action is strengthened.