scholarly journals Identification of the epitope for a monoclonal antibody that blocks platelet aggregation induced by type III collagen

1997 ◽  
Vol 323 (1) ◽  
pp. 45-49 ◽  
Author(s):  
Veronica GLATTAUER ◽  
Jerome A. WERKMEISTER ◽  
Alan KIRKPATRICK ◽  
John A. M. RAMSHAW
Keyword(s):  
Monoclonal Antibody ◽  
Platelet Aggregation ◽  
Type Iii Collagen ◽  
Inhibit Platelet Aggregation ◽  
Inhibitory Antibody ◽  
Type Iii ◽  
Human Type ◽  
Letter Code ◽  
The One ◽  
Α2β1 Integrin

A library of eight conformation-dependent monoclonal antibodies that react with distinct epitopes on native human type III collagen has been examined for the ability of these antibodies to inhibit platelet aggregation induced by this collagen. Six of these antibodies had no effects; one, 1E7-D7/Col3, delayed the onset and slowed the rate of platelet aggregation, while another, 2G8-B1/Col3, completely inhibited aggregation. In order to identify the epitope recognized by this inhibitory antibody, a series of peptides that could fold to form triple-helical fragments was examined. Each peptide included six Gly-Xaa-Yaa triplets from the human type III collagen sequence, where Xaa and Yaa represent the particular amino acids in the sequence, and a C-terminal (Gly-Pro-Hyp)4 sequence to enhance triple-helical stability. Using these peptides we have identified the epitope as a nine-amino-acid sequence, GLAGAOGLR (where O is the one-letter code for 4-hydroxyproline), starting at position 520 in the human type III collagen helical domain. This sequence is proximal to the site proposed for the interaction of type III collagen with α2β1-integrin of platelets.

scholarly journals Production and characterization of a monoclonal antibody to human type III collagen.

1986 ◽  
Vol 34 (8) ◽  
pp. 1003-1011 ◽  
Author(s):  
E J Macarak ◽  
P S Howard ◽  
E T Lally
Keyword(s):  
Monoclonal Antibody ◽  
Lymph Node ◽  
Staining Pattern ◽  
Immunohistochemical Localization ◽  
White Pulp ◽  
Type Iii Collagen ◽  
Type I ◽  
Type Iii ◽  
Human Type ◽  
New Information

Human type III collagen from placenta was isolated and purified for use as an immunogen. A monoclonal antibody was produced which specifically recognizes epitopes unique to type III collagen. The specificity of the antibody was determined by inhibition ELISA, an immunoblot assay, and by immunoprecipitation. Results indicated that the monoclonal antibody recognized only the alpha 1(III) polypeptide chains and did not crossreact with type I, IV, or V collagen. The monoclonal antibody was also used for immunohistochemical localization of type III collagen in tissue sections of human placenta, bovine spleen, and lymph node. In placenta, both large and small blood vessels showed pronounced staining of the tunica media, which contains largely smooth muscle cells, known to synthesize type III collagen. In contrast, the intimal areas and endothelial cells showed no staining with the antibody. In the placental villi, staining was limited to the villous core, where fine fibrillar structures showed strong staining. In lymph nodes, the capsule and pericapsular adipose cells were surrounded by a covering of type III collagen. Within the parenchyma of the node, staining was localized to a branching, reticular array of fine fibers. In the spleen, staining was pronounced in the capsule, splenic trabeculae, and white pulp, where blood vessel staining was especially prominent. The red pulp and splenic sinuses contain little or no type III collagen. The fine network-like or reticular staining pattern found in the lymph node parenchyma is consistent with the staining pattern of the protein reticulin, and suggests that type III collagen may be closely associated with reticulin in certain tissues. Since the role of type III in tissues is unclear, this reagent will be useful in providing new information in this regard.

Inhibition by Peptidic Fragments of Clq of Platelet - Collagen Interactions

1979 ◽  
Author(s):  
J.L. wautier ◽  
K.B.M. Reid ◽  
Y. Legrand ◽  
J.P. Caen
Keyword(s):  
Amino Acid ◽  
Platelet Aggregation ◽  
Amino Acid Sequence ◽  
Platelet Adhesion ◽  
Type Iii Collagen ◽  
Type I ◽  
Inhibit Platelet Aggregation ◽  
Pepsin Digestion ◽  
Type Iii ◽  
Collagen Peptides

Clq (first component of complement) has been shown to inhibit platelet aggregation induced by collagen. Platelet aggregation and adhesion to purified type I or type III collagen have been studied in the presence of various fragment of Clq.The collagen like region was obtained by digestion of Clq by pepsin and by isolation of the fragments by chromatography on sephadex G 200. The globular region was prepared by digestion of Clq by collagenase, and filtration. The collagen like region is as effective as native Clq in inhibiting platelet aggregation or adhesion in the presence of type I or type III collagen, while the globular region was without effect.Using peptides obtained from the collagen like region of Clq (reduction and alkylation, performic oxidation, CNBr treatment, extensive pepsin digestion) it could be shown that the fragment (FR II = 70 Amino Acid) was responsible for the inhibition of the platelet adhesion and aggregation induced by type I or type III collagen.The Amino Acid sequence of this fragment was compared to the known sequence of the collagen peptides involved in platelet adhesion (see Fauvel et al).

scholarly journals Characterization of Human Type III Collagen Expressed in a Baculovirus System

1996 ◽  
Vol 271 (20) ◽  
pp. 11988-11995 ◽  
Author(s):  
Arja Lamberg ◽  
Tarja Helaakoski ◽  
Johanna Myllyharju ◽  
Sirkku Peltonen ◽  
Holger Notbohm ◽  
...  
Keyword(s):  
Type Iii Collagen ◽  
Type Iii ◽  
Human Type ◽  
Baculovirus System

scholarly journals Crystal Structure of Human Type III Collagen Gly991–Gly1032Cystine Knot-containing Peptide Shows Both 7/2 and 10/3 Triple Helical Symmetries

2008 ◽  
Vol 283 (47) ◽  
pp. 32580-32589 ◽  
Author(s):  
Sergei P. Boudko ◽  
Jürgen Engel ◽  
Kenji Okuyama ◽  
Kazunori Mizuno ◽  
Hans Peter Bächinger ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Type Iii Collagen ◽  
Type Iii ◽  
Human Type

scholarly journals An Ehrlich chromogen in collagen cross-links

1983 ◽  
Vol 209 (1) ◽  
pp. 263-264 ◽  
Author(s):  
J E Scott ◽  
R G Qian ◽  
W Henkel ◽  
R W Glanville
Keyword(s):  
Type Iv Collagen ◽  
Rich Source ◽  
Type Iii Collagen ◽  
Cross Link ◽  
Type Iii ◽  
Human Type ◽  
Type Iv ◽  
Cross Links

A well-characterized three-chain peptide [(Col1)2 X T9] from human type III collagen was a rich source of Ehrlich chromogen. The corresponding two-chain peptide [(Col1)2] was not, implying that the Ehrlich chromogen is a trifunctional cross-link. (Col1)2 X T9 also contained pyridinoline, which is not an Ehrlich chromogen. The 7S domain of type IV collagen also contained an Ehrlich chromogen.

scholarly journals Histological evaluation of the periodontal ligament from aged wistar rats supplemented with ascorbic acid

2013 ◽  
Vol 85 (1) ◽  
pp. 327-335 ◽  
Author(s):  
Jacqueline N. Zanoni ◽  
Nathalia M. Lucas ◽  
Aline R. Trevizan ◽  
Ivan D.S. Souza
Keyword(s):  
Ascorbic Acid ◽  
Periodontal Ligament ◽  
Aging Process ◽  
Natural Aging ◽  
Collagen Fibers ◽  
Histological Evaluation ◽  
Type Iii Collagen ◽  
Type I ◽  
Type Iii ◽  
The One

Ascorbic acid (AA) is able to neutralize reactive oxygen species and is essential for collagen synthesis. In aging process oxidative stress is elevated. This study aims to investigate the effects of AA supplementation on the periodontal ligament (PL) of rats during aging. Twenty five rats were used and divided into groups: J90 (90-day-old control), E345 (345-day-old control), E428 (428-day-old control), EA345 (345-day-old supplemented with AA from 90-day-old on) and EA428 (428-day-old supplemented with AA from 90-day-old on). We analyzed the thickness, density of fibroblasts and blood vessels and collagen fibers types in the PL. In group J90 there was predominantly type III collagen fibers (87.64%). In animals supplemented with AA, the area filled by type I fibers (group EA345: 65.67%, group EA428: 52.23%) was higher than type III fibers. PL in group EA428 was thicker than the one observed in group E428 (P < 0.05). During natural aging process, AA promoted the maturation of collagen fibers and enhanced angiogenesis in periodontal ligament. One can conclude that the supplementation with AA represented a beneficial factor for the development of PL in aged rats.

IGM antibodies can be used as direct immunoelectron markers

1987 ◽  
Vol 45 ◽  
pp. 764-765
Author(s):  
Douglas R. Keene ◽  
Lynn Y. Sakai ◽  
Robert E. Burgeson ◽  
Hans Peter Bächinger
Keyword(s):  
Electron Density ◽  
Colloidal Gold ◽  
Type Iii Collagen ◽  
Secondary Antibody ◽  
Type Iii ◽  
Human Type ◽  
Igm Antibodies ◽  
Electron Microscopes ◽  
Scanning Electron Microscopes ◽  
Scanning Electron

A mouse monoclonal IgM antibody has been used in this study as a direct ultrastructural marker to immunolocalize human Type III collagen at the level of the transmission and scanning electron microscopes. The method utilized involves the use of none of the electron dense conjugates such as colloidal gold, ferritin, or peroxidase: Hence ultrastructural immuno- localization relies exclusively on the size and electron density of the bound IgM complex after routine staining. As compared to similar immuno- localization studies utilizing the same monoclonal anti-Type III collagen IgM but with additional secondary antibody-gold conjugates, the direct “naked” IgM technique consistently gave results of higher specificity and greater sensitivity.

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