Loading…

Thrombin and fibrinogen γ′ impact clot structure by marked effects on intrafibrillar structure and protofibril packing

Previous studies have shown effects of thrombin and fibrinogen γ′ on clot structure. However, structural information was obtained using electron microscopy, which requires sample dehydration. Our aim was to investigate the role of thrombin and fibrinogen γ′ in modulating fibrin structure under fully...

Full description

Saved in:
Bibliographic Details
Published in:Blood 2016-01, Vol.127 (4), p.487-495
Main Authors: Domingues, Marco M., Macrae, Fraser L., Duval, Cédric, McPherson, Helen R., Bridge, Katherine I., Ajjan, Ramzi A., Ridger, Victoria C., Connell, Simon D., Philippou, Helen, Ariëns, Robert A.S.
Format: Article
Language:English
Subjects:
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-c408t-f02ab6afbb7c58f6436d765455ee83318ee8268af971e3af78f34adcf54daa193
cites cdi_FETCH-LOGICAL-c408t-f02ab6afbb7c58f6436d765455ee83318ee8268af971e3af78f34adcf54daa193
container_end_page 495
container_issue 4
container_start_page 487
container_title Blood
container_volume 127
creator Domingues, Marco M.
Macrae, Fraser L.
Duval, Cédric
McPherson, Helen R.
Bridge, Katherine I.
Ajjan, Ramzi A.
Ridger, Victoria C.
Connell, Simon D.
Philippou, Helen
Ariëns, Robert A.S.
description Previous studies have shown effects of thrombin and fibrinogen γ′ on clot structure. However, structural information was obtained using electron microscopy, which requires sample dehydration. Our aim was to investigate the role of thrombin and fibrinogen γ′ in modulating fibrin structure under fully hydrated conditions. Fibrin fibers were studied using turbidimetry, atomic force microscopy, electron microscopy, and magnetic tweezers in purified and plasma solutions. Increased thrombin induced a pronounced decrease in average protofibril content per fiber, with a relatively minor decrease in fiber size, leading to the formation of less compact fiber structures. Atomic force microscopy under fully hydrated conditions confirmed that fiber diameter was only marginally decreased. Decreased protofibril content of the fibers produced by high thrombin resulted in weakened clot architecture as analyzed by magnetic tweezers in purified systems and by thromboelastometry in plasma and whole blood. Fibers produced with fibrinogen γ′ showed reduced protofibril packing over a range of thrombin concentrations. High-magnification electron microscopy demonstrated reduced protofibril packing in γ′ fibers and unraveling of fibers into separate protofibrils. Decreased protofibril packing was confirmed in plasma for high thrombin concentrations and fibrinogen-deficient plasma reconstituted with γ′ fibrinogen. These findings demonstrate that, in fully hydrated conditions, thrombin and fibrinogen γ′ have dramatic effects on protofibril content and that protein density within fibers correlates with strength of the fibrin network. We conclude that regulation of protofibril content of fibers is an important mechanism by which thrombin and fibrinogen γ′ modulate fibrin clot structure and strength. •Thrombin and fibrinogen γ′ regulate protofibril packing within fibrin fibers and thereby influence clot stiffness.•Fibrin analysis after dehydration (e.g. electron microscopy) overestimates changes in fiber size due to effects on protofibril packing.
doi_str_mv 10.1182/blood-2015-06-652214
format article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1761472871</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S000649712030522X</els_id><sourcerecordid>1761472871</sourcerecordid><originalsourceid>FETCH-LOGICAL-c408t-f02ab6afbb7c58f6436d765455ee83318ee8268af971e3af78f34adcf54daa193</originalsourceid><addsrcrecordid>eNp9kE1uFDEQhS1ERIbADRDykk0H2227PRskFIUfKRKbZG257XIw6bYH2400u5wJ7sEhOEk86YBYZVWLeu9VvQ-hV5ScUqrY23FKyXWMUNER2UnBGOVP0IYKpjpCGHmKNoS0Dd8O9Bg9L-UbIZT3TDxDx0xKonq23aD95dec5jFEbKLDPow5xHQNEf_-9ef2Jw7zztiK7ZQqLjUvti4Z8LjHs8k34DB4D7YWnCIOsWZzHzBNJv-nPgTvcqppXeKWeBPi9Qt05M1U4OXDPEFXH84vzz51F18-fj57f9FZTlTtPGFmlMaP42CF8pL30g1ScCEAVN9T1QaTyvhWE3rjB-V7bpz1gjtj6LY_QW_W3PbD9wVK1XMoFtqTEdJSNB0k5QNTA21SvkptTqVk8HqXQ2u615ToA3R9D10foGsi9Qq92V4_XFjGGdw_01_KTfBuFUDr-SNA1sUGiBZcyA2fdik8fuEO8dmYJA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1761472871</pqid></control><display><type>article</type><title>Thrombin and fibrinogen γ′ impact clot structure by marked effects on intrafibrillar structure and protofibril packing</title><source>ScienceDirect (Online service)</source><creator>Domingues, Marco M. ; Macrae, Fraser L. ; Duval, Cédric ; McPherson, Helen R. ; Bridge, Katherine I. ; Ajjan, Ramzi A. ; Ridger, Victoria C. ; Connell, Simon D. ; Philippou, Helen ; Ariëns, Robert A.S.</creator><creatorcontrib>Domingues, Marco M. ; Macrae, Fraser L. ; Duval, Cédric ; McPherson, Helen R. ; Bridge, Katherine I. ; Ajjan, Ramzi A. ; Ridger, Victoria C. ; Connell, Simon D. ; Philippou, Helen ; Ariëns, Robert A.S.</creatorcontrib><description>Previous studies have shown effects of thrombin and fibrinogen γ′ on clot structure. However, structural information was obtained using electron microscopy, which requires sample dehydration. Our aim was to investigate the role of thrombin and fibrinogen γ′ in modulating fibrin structure under fully hydrated conditions. Fibrin fibers were studied using turbidimetry, atomic force microscopy, electron microscopy, and magnetic tweezers in purified and plasma solutions. Increased thrombin induced a pronounced decrease in average protofibril content per fiber, with a relatively minor decrease in fiber size, leading to the formation of less compact fiber structures. Atomic force microscopy under fully hydrated conditions confirmed that fiber diameter was only marginally decreased. Decreased protofibril content of the fibers produced by high thrombin resulted in weakened clot architecture as analyzed by magnetic tweezers in purified systems and by thromboelastometry in plasma and whole blood. Fibers produced with fibrinogen γ′ showed reduced protofibril packing over a range of thrombin concentrations. High-magnification electron microscopy demonstrated reduced protofibril packing in γ′ fibers and unraveling of fibers into separate protofibrils. Decreased protofibril packing was confirmed in plasma for high thrombin concentrations and fibrinogen-deficient plasma reconstituted with γ′ fibrinogen. These findings demonstrate that, in fully hydrated conditions, thrombin and fibrinogen γ′ have dramatic effects on protofibril content and that protein density within fibers correlates with strength of the fibrin network. We conclude that regulation of protofibril content of fibers is an important mechanism by which thrombin and fibrinogen γ′ modulate fibrin clot structure and strength. •Thrombin and fibrinogen γ′ regulate protofibril packing within fibrin fibers and thereby influence clot stiffness.•Fibrin analysis after dehydration (e.g. electron microscopy) overestimates changes in fiber size due to effects on protofibril packing.</description><identifier>ISSN: 0006-4971</identifier><identifier>EISSN: 1528-0020</identifier><identifier>DOI: 10.1182/blood-2015-06-652214</identifier><identifier>PMID: 26608329</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Blood Coagulation ; Blood Viscosity ; Fibrinogens, Abnormal - metabolism ; Fibrinogens, Abnormal - ultrastructure ; Humans ; Microscopy, Atomic Force ; Nephelometry and Turbidimetry ; Thrombin - metabolism ; Thrombin - ultrastructure ; Thrombosis - metabolism</subject><ispartof>Blood, 2016-01, Vol.127 (4), p.487-495</ispartof><rights>2016 American Society of Hematology</rights><rights>2016 by The American Society of Hematology.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c408t-f02ab6afbb7c58f6436d765455ee83318ee8268af971e3af78f34adcf54daa193</citedby><cites>FETCH-LOGICAL-c408t-f02ab6afbb7c58f6436d765455ee83318ee8268af971e3af78f34adcf54daa193</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S000649712030522X$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,780,784,3549,27924,27925,45780</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26608329$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Domingues, Marco M.</creatorcontrib><creatorcontrib>Macrae, Fraser L.</creatorcontrib><creatorcontrib>Duval, Cédric</creatorcontrib><creatorcontrib>McPherson, Helen R.</creatorcontrib><creatorcontrib>Bridge, Katherine I.</creatorcontrib><creatorcontrib>Ajjan, Ramzi A.</creatorcontrib><creatorcontrib>Ridger, Victoria C.</creatorcontrib><creatorcontrib>Connell, Simon D.</creatorcontrib><creatorcontrib>Philippou, Helen</creatorcontrib><creatorcontrib>Ariëns, Robert A.S.</creatorcontrib><title>Thrombin and fibrinogen γ′ impact clot structure by marked effects on intrafibrillar structure and protofibril packing</title><title>Blood</title><addtitle>Blood</addtitle><description>Previous studies have shown effects of thrombin and fibrinogen γ′ on clot structure. However, structural information was obtained using electron microscopy, which requires sample dehydration. Our aim was to investigate the role of thrombin and fibrinogen γ′ in modulating fibrin structure under fully hydrated conditions. Fibrin fibers were studied using turbidimetry, atomic force microscopy, electron microscopy, and magnetic tweezers in purified and plasma solutions. Increased thrombin induced a pronounced decrease in average protofibril content per fiber, with a relatively minor decrease in fiber size, leading to the formation of less compact fiber structures. Atomic force microscopy under fully hydrated conditions confirmed that fiber diameter was only marginally decreased. Decreased protofibril content of the fibers produced by high thrombin resulted in weakened clot architecture as analyzed by magnetic tweezers in purified systems and by thromboelastometry in plasma and whole blood. Fibers produced with fibrinogen γ′ showed reduced protofibril packing over a range of thrombin concentrations. High-magnification electron microscopy demonstrated reduced protofibril packing in γ′ fibers and unraveling of fibers into separate protofibrils. Decreased protofibril packing was confirmed in plasma for high thrombin concentrations and fibrinogen-deficient plasma reconstituted with γ′ fibrinogen. These findings demonstrate that, in fully hydrated conditions, thrombin and fibrinogen γ′ have dramatic effects on protofibril content and that protein density within fibers correlates with strength of the fibrin network. We conclude that regulation of protofibril content of fibers is an important mechanism by which thrombin and fibrinogen γ′ modulate fibrin clot structure and strength. •Thrombin and fibrinogen γ′ regulate protofibril packing within fibrin fibers and thereby influence clot stiffness.•Fibrin analysis after dehydration (e.g. electron microscopy) overestimates changes in fiber size due to effects on protofibril packing.</description><subject>Blood Coagulation</subject><subject>Blood Viscosity</subject><subject>Fibrinogens, Abnormal - metabolism</subject><subject>Fibrinogens, Abnormal - ultrastructure</subject><subject>Humans</subject><subject>Microscopy, Atomic Force</subject><subject>Nephelometry and Turbidimetry</subject><subject>Thrombin - metabolism</subject><subject>Thrombin - ultrastructure</subject><subject>Thrombosis - metabolism</subject><issn>0006-4971</issn><issn>1528-0020</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><recordid>eNp9kE1uFDEQhS1ERIbADRDykk0H2227PRskFIUfKRKbZG257XIw6bYH2400u5wJ7sEhOEk86YBYZVWLeu9VvQ-hV5ScUqrY23FKyXWMUNER2UnBGOVP0IYKpjpCGHmKNoS0Dd8O9Bg9L-UbIZT3TDxDx0xKonq23aD95dec5jFEbKLDPow5xHQNEf_-9ef2Jw7zztiK7ZQqLjUvti4Z8LjHs8k34DB4D7YWnCIOsWZzHzBNJv-nPgTvcqppXeKWeBPi9Qt05M1U4OXDPEFXH84vzz51F18-fj57f9FZTlTtPGFmlMaP42CF8pL30g1ScCEAVN9T1QaTyvhWE3rjB-V7bpz1gjtj6LY_QW_W3PbD9wVK1XMoFtqTEdJSNB0k5QNTA21SvkptTqVk8HqXQ2u615ToA3R9D10foGsi9Qq92V4_XFjGGdw_01_KTfBuFUDr-SNA1sUGiBZcyA2fdik8fuEO8dmYJA</recordid><startdate>20160128</startdate><enddate>20160128</enddate><creator>Domingues, Marco M.</creator><creator>Macrae, Fraser L.</creator><creator>Duval, Cédric</creator><creator>McPherson, Helen R.</creator><creator>Bridge, Katherine I.</creator><creator>Ajjan, Ramzi A.</creator><creator>Ridger, Victoria C.</creator><creator>Connell, Simon D.</creator><creator>Philippou, Helen</creator><creator>Ariëns, Robert A.S.</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20160128</creationdate><title>Thrombin and fibrinogen γ′ impact clot structure by marked effects on intrafibrillar structure and protofibril packing</title><author>Domingues, Marco M. ; Macrae, Fraser L. ; Duval, Cédric ; McPherson, Helen R. ; Bridge, Katherine I. ; Ajjan, Ramzi A. ; Ridger, Victoria C. ; Connell, Simon D. ; Philippou, Helen ; Ariëns, Robert A.S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c408t-f02ab6afbb7c58f6436d765455ee83318ee8268af971e3af78f34adcf54daa193</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Blood Coagulation</topic><topic>Blood Viscosity</topic><topic>Fibrinogens, Abnormal - metabolism</topic><topic>Fibrinogens, Abnormal - ultrastructure</topic><topic>Humans</topic><topic>Microscopy, Atomic Force</topic><topic>Nephelometry and Turbidimetry</topic><topic>Thrombin - metabolism</topic><topic>Thrombin - ultrastructure</topic><topic>Thrombosis - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Domingues, Marco M.</creatorcontrib><creatorcontrib>Macrae, Fraser L.</creatorcontrib><creatorcontrib>Duval, Cédric</creatorcontrib><creatorcontrib>McPherson, Helen R.</creatorcontrib><creatorcontrib>Bridge, Katherine I.</creatorcontrib><creatorcontrib>Ajjan, Ramzi A.</creatorcontrib><creatorcontrib>Ridger, Victoria C.</creatorcontrib><creatorcontrib>Connell, Simon D.</creatorcontrib><creatorcontrib>Philippou, Helen</creatorcontrib><creatorcontrib>Ariëns, Robert A.S.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Blood</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Domingues, Marco M.</au><au>Macrae, Fraser L.</au><au>Duval, Cédric</au><au>McPherson, Helen R.</au><au>Bridge, Katherine I.</au><au>Ajjan, Ramzi A.</au><au>Ridger, Victoria C.</au><au>Connell, Simon D.</au><au>Philippou, Helen</au><au>Ariëns, Robert A.S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Thrombin and fibrinogen γ′ impact clot structure by marked effects on intrafibrillar structure and protofibril packing</atitle><jtitle>Blood</jtitle><addtitle>Blood</addtitle><date>2016-01-28</date><risdate>2016</risdate><volume>127</volume><issue>4</issue><spage>487</spage><epage>495</epage><pages>487-495</pages><issn>0006-4971</issn><eissn>1528-0020</eissn><abstract>Previous studies have shown effects of thrombin and fibrinogen γ′ on clot structure. However, structural information was obtained using electron microscopy, which requires sample dehydration. Our aim was to investigate the role of thrombin and fibrinogen γ′ in modulating fibrin structure under fully hydrated conditions. Fibrin fibers were studied using turbidimetry, atomic force microscopy, electron microscopy, and magnetic tweezers in purified and plasma solutions. Increased thrombin induced a pronounced decrease in average protofibril content per fiber, with a relatively minor decrease in fiber size, leading to the formation of less compact fiber structures. Atomic force microscopy under fully hydrated conditions confirmed that fiber diameter was only marginally decreased. Decreased protofibril content of the fibers produced by high thrombin resulted in weakened clot architecture as analyzed by magnetic tweezers in purified systems and by thromboelastometry in plasma and whole blood. Fibers produced with fibrinogen γ′ showed reduced protofibril packing over a range of thrombin concentrations. High-magnification electron microscopy demonstrated reduced protofibril packing in γ′ fibers and unraveling of fibers into separate protofibrils. Decreased protofibril packing was confirmed in plasma for high thrombin concentrations and fibrinogen-deficient plasma reconstituted with γ′ fibrinogen. These findings demonstrate that, in fully hydrated conditions, thrombin and fibrinogen γ′ have dramatic effects on protofibril content and that protein density within fibers correlates with strength of the fibrin network. We conclude that regulation of protofibril content of fibers is an important mechanism by which thrombin and fibrinogen γ′ modulate fibrin clot structure and strength. •Thrombin and fibrinogen γ′ regulate protofibril packing within fibrin fibers and thereby influence clot stiffness.•Fibrin analysis after dehydration (e.g. electron microscopy) overestimates changes in fiber size due to effects on protofibril packing.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>26608329</pmid><doi>10.1182/blood-2015-06-652214</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 0006-4971
ispartof Blood, 2016-01, Vol.127 (4), p.487-495
issn 0006-4971
1528-0020
language eng
recordid cdi_proquest_miscellaneous_1761472871
source ScienceDirect (Online service)
subjects Blood Coagulation
Blood Viscosity
Fibrinogens, Abnormal - metabolism
Fibrinogens, Abnormal - ultrastructure
Humans
Microscopy, Atomic Force
Nephelometry and Turbidimetry
Thrombin - metabolism
Thrombin - ultrastructure
Thrombosis - metabolism
title Thrombin and fibrinogen γ′ impact clot structure by marked effects on intrafibrillar structure and protofibril packing
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T14%3A30%3A39IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Thrombin%20and%20fibrinogen%20%CE%B3%E2%80%B2%20impact%20clot%20structure%20by%20marked%20effects%20on%20intrafibrillar%20structure%20and%20protofibril%20packing&rft.jtitle=Blood&rft.au=Domingues,%20Marco%20M.&rft.date=2016-01-28&rft.volume=127&rft.issue=4&rft.spage=487&rft.epage=495&rft.pages=487-495&rft.issn=0006-4971&rft.eissn=1528-0020&rft_id=info:doi/10.1182/blood-2015-06-652214&rft_dat=%3Cproquest_cross%3E1761472871%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c408t-f02ab6afbb7c58f6436d765455ee83318ee8268af971e3af78f34adcf54daa193%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=1761472871&rft_id=info:pmid/26608329&rfr_iscdi=true