Structural and functional studies of novel viral complement inhibitors

Linda Mark

Structural and functional studies of novel viral complement inhibitors

Linda Mark

Clinical Chemistry, Malmö

Research output: Thesis › Doctoral Thesis (compilation)

Abstract

Viruses have developed several strategies to evade the complement system, a part of the immune system. The fourth open reading frame of the human virus Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a type 1 transmembrane protein. This contains four complement control protein domains (CCP domains), and is thus structurally similar to human complement inhibitors. We named it KSHV complement control protein (KCP). Using recombinant KCP, we showed that it can inhibit the complement system by disrupting a key enzymatic complex, the C3 convertase. We predicted the 3D structure of KCP by homology modeling. Functionally important sites in KCP were mapped using mutants where either one to three amino acids, or entire CCP domains, were altered.

We also found that a patch of positively charged amino acids in CCP 1-2 of KCP can bind to heparin and weakly to heparin like proteoglycans at the surface of cells, which seems to aid the virus in the attachment step prior to cell entry.

In order to study the biological role of complement inhibition for this type of virus, we want to use the KSHV homologue rhesus rhadinovirus (RRV), since this virus is both technically easier to work with than KCP and also permits in vivo studies, by naturally infecting Rhesus Macaques. RRV complement control protein (RCP) is the KCP homologue in RRV. We have characterized its gene, confirmed that RCP is expressed and present on the surface of virus and performed experiments indicating that it is a functional complement inhibitor.

Original language	English
Qualification	Doctor
Awarding Institution	Clinical Chemistry, Malmö
Supervisors/Advisors	Blom, Anna, Supervisor
Award date	2006 Dec 8
Publisher	Department of Laboratory Medicine, Lund University
ISBN (Print)	91-85559-62-8
Publication status	Published - 2006

Bibliographical note

Defence details

Date: 2006-12-08
Time: 09:15
Place: Lilla aulan, ing 59 ,UMAS, Malmö

External reviewer(s)

Name: Rodriguez de Cordoba, Santiago
Title: Dr
Affiliation: Departamento de Inmunologia,

---

<div class="article_info">O.B. Spiller, D.J. Blackbourn, L Mark, D Proctor and A.M. Blom. <span class="article_issue_date">2003</span>. <span class="article_title">Functional Activity of the Complement Regulator Encoded by Kaposi's Sarcoma-associated Herpesvirus.</span> <span class="journal_series_title">The J Biol Chem</span>, <span class="journal_volume">vol 278</span> <span class="journal_pages">pp 9283-9289</span>.</div>
<div class="article_info">L Mark, WH Lee, OB Spiller, D Proctor, DJ Blackbourn, BO Villoutreix and AM Blom. <span class="article_issue_date">2004</span>. <span class="article_title">The Kaposi's Sarcoma-associated Herpesvirus Complement Control Protein Mimics Human Molecular Mechanisms for Inhibition of the Complement System.</span> <span class="journal_series_title">J Biol Chem</span>, <span class="journal_volume">vol 279</span> <span class="journal_pages">pp 45093-101</span>.</div>
<div class="article_info">L Mark, WH Lee, OB Spiller, BO Villoutreix and AM Blom. <span class="article_issue_date">2006</span>. <span class="article_title">The Kaposi's sarcoma-associated herpesvirus complement control protein (KCP) binds to heparin and cell surfaces via positively charged amino acids in CCP1-2.</span> <span class="journal_series_title">Mol Immunol</span>, <span class="journal_volume">vol 43</span> <span class="journal_pages">pp 1665-75</span>.</div>
<div class="article_info">OB Spiller, L Mark, CE Blue, DG Proctor, JA Aitken, AM Blom and DJ Blackbourn. <span class="article_issue_date">2006</span>. <span class="article_title">Dissecting the regions of virion-associated Kaposi's sarcoma-associated herpesvirus complement control protein required for complement regulation and cell binding.</span> <span class="journal_series_title">J Virol</span>, <span class="journal_volume">vol 80</span> <span class="journal_pages">pp 4068-78</span>.</div>
<div class="article_info">L Mark, OB Spiller, M Okroj, S Chanas, JA Aitken, SW Wong, B Damania, AM Blom and DJ Blackbourn. <span class="article_issue_date"></span>. <span class="article_title">Molecular characterization of the Rhesus Rhadinovirus (RRV) ORF4 gene and the complement control protein (RCP) it encodes.</span> (submitted)</div>

Subject classification (UKÄ)

Medicinal Chemistry

Free keywords

transplantation
serologi
serology
Kaposi's sarcoma-associated herpesvirus
Immunologi
Immunology
homology modeling
complement inhibitor

Cite this

@phdthesis{549b460394094662a27395a9018af58a,

title = "Structural and functional studies of novel viral complement inhibitors",

abstract = "Viruses have developed several strategies to evade the complement system, a part of the immune system. The fourth open reading frame of the human virus Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a type 1 transmembrane protein. This contains four complement control protein domains (CCP domains), and is thus structurally similar to human complement inhibitors. We named it KSHV complement control protein (KCP). Using recombinant KCP, we showed that it can inhibit the complement system by disrupting a key enzymatic complex, the C3 convertase. We predicted the 3D structure of KCP by homology modeling. Functionally important sites in KCP were mapped using mutants where either one to three amino acids, or entire CCP domains, were altered. We also found that a patch of positively charged amino acids in CCP 1-2 of KCP can bind to heparin and weakly to heparin like proteoglycans at the surface of cells, which seems to aid the virus in the attachment step prior to cell entry. In order to study the biological role of complement inhibition for this type of virus, we want to use the KSHV homologue rhesus rhadinovirus (RRV), since this virus is both technically easier to work with than KCP and also permits in vivo studies, by naturally infecting Rhesus Macaques. RRV complement control protein (RCP) is the KCP homologue in RRV. We have characterized its gene, confirmed that RCP is expressed and present on the surface of virus and performed experiments indicating that it is a functional complement inhibitor.",

keywords = "transplantation, serologi, serology, Kaposi's sarcoma-associated herpesvirus, Immunologi, Immunology, homology modeling, complement inhibitor",

author = "Linda Mark",

note = "Defence details Date: 2006-12-08 Time: 09:15 Place: Lilla aulan, ing 59 ,UMAS, Malm{\"o} External reviewer(s) Name: Rodriguez de Cordoba, Santiago Title: Dr Affiliation: Departamento de Inmunologia, --- <div class={"}article_info{"}>O.B. Spiller, D.J. Blackbourn, L Mark, D Proctor and A.M. Blom. <span class={"}article_issue_date{"}>2003</span>. <span class={"}article_title{"}>Functional Activity of the Complement Regulator Encoded by Kaposi's Sarcoma-associated Herpesvirus.</span> <span class={"}journal_series_title{"}>The J Biol Chem</span>, <span class={"}journal_volume{"}>vol 278</span> <span class={"}journal_pages{"}>pp 9283-9289</span>.</div> <div class={"}article_info{"}>L Mark, WH Lee, OB Spiller, D Proctor, DJ Blackbourn, BO Villoutreix and AM Blom. <span class={"}article_issue_date{"}>2004</span>. <span class={"}article_title{"}>The Kaposi's Sarcoma-associated Herpesvirus Complement Control Protein Mimics Human Molecular Mechanisms for Inhibition of the Complement System.</span> <span class={"}journal_series_title{"}>J Biol Chem</span>, <span class={"}journal_volume{"}>vol 279</span> <span class={"}journal_pages{"}>pp 45093-101</span>.</div> <div class={"}article_info{"}>L Mark, WH Lee, OB Spiller, BO Villoutreix and AM Blom. <span class={"}article_issue_date{"}>2006</span>. <span class={"}article_title{"}>The Kaposi's sarcoma-associated herpesvirus complement control protein (KCP) binds to heparin and cell surfaces via positively charged amino acids in CCP1-2.</span> <span class={"}journal_series_title{"}>Mol Immunol</span>, <span class={"}journal_volume{"}>vol 43</span> <span class={"}journal_pages{"}>pp 1665-75</span>.</div> <div class={"}article_info{"}>OB Spiller, L Mark, CE Blue, DG Proctor, JA Aitken, AM Blom and DJ Blackbourn. <span class={"}article_issue_date{"}>2006</span>. <span class={"}article_title{"}>Dissecting the regions of virion-associated Kaposi's sarcoma-associated herpesvirus complement control protein required for complement regulation and cell binding.</span> <span class={"}journal_series_title{"}>J Virol</span>, <span class={"}journal_volume{"}>vol 80</span> <span class={"}journal_pages{"}>pp 4068-78</span>.</div> <div class={"}article_info{"}>L Mark, OB Spiller, M Okroj, S Chanas, JA Aitken, SW Wong, B Damania, AM Blom and DJ Blackbourn. <span class={"}article_issue_date{"}></span>. <span class={"}article_title{"}>Molecular characterization of the Rhesus Rhadinovirus (RRV) ORF4 gene and the complement control protein (RCP) it encodes.</span> (submitted)</div>",

year = "2006",

language = "English",

isbn = "91-85559-62-8",

series = "2006:140",

publisher = "Department of Laboratory Medicine, Lund University",

type = "Doctoral Thesis (compilation)",

school = "Clinical Chemistry, Malm{\"o}",

}

TY - THES

T1 - Structural and functional studies of novel viral complement inhibitors

AU - Mark, Linda

N1 - Defence details Date: 2006-12-08 Time: 09:15 Place: Lilla aulan, ing 59 ,UMAS, Malmö External reviewer(s) Name: Rodriguez de Cordoba, Santiago Title: Dr Affiliation: Departamento de Inmunologia, --- <div class="article_info">O.B. Spiller, D.J. Blackbourn, L Mark, D Proctor and A.M. Blom. <span class="article_issue_date">2003</span>. <span class="article_title">Functional Activity of the Complement Regulator Encoded by Kaposi's Sarcoma-associated Herpesvirus.</span> <span class="journal_series_title">The J Biol Chem</span>, <span class="journal_volume">vol 278</span> <span class="journal_pages">pp 9283-9289</span>.</div> <div class="article_info">L Mark, WH Lee, OB Spiller, D Proctor, DJ Blackbourn, BO Villoutreix and AM Blom. <span class="article_issue_date">2004</span>. <span class="article_title">The Kaposi's Sarcoma-associated Herpesvirus Complement Control Protein Mimics Human Molecular Mechanisms for Inhibition of the Complement System.</span> <span class="journal_series_title">J Biol Chem</span>, <span class="journal_volume">vol 279</span> <span class="journal_pages">pp 45093-101</span>.</div> <div class="article_info">L Mark, WH Lee, OB Spiller, BO Villoutreix and AM Blom. <span class="article_issue_date">2006</span>. <span class="article_title">The Kaposi's sarcoma-associated herpesvirus complement control protein (KCP) binds to heparin and cell surfaces via positively charged amino acids in CCP1-2.</span> <span class="journal_series_title">Mol Immunol</span>, <span class="journal_volume">vol 43</span> <span class="journal_pages">pp 1665-75</span>.</div> <div class="article_info">OB Spiller, L Mark, CE Blue, DG Proctor, JA Aitken, AM Blom and DJ Blackbourn. <span class="article_issue_date">2006</span>. <span class="article_title">Dissecting the regions of virion-associated Kaposi's sarcoma-associated herpesvirus complement control protein required for complement regulation and cell binding.</span> <span class="journal_series_title">J Virol</span>, <span class="journal_volume">vol 80</span> <span class="journal_pages">pp 4068-78</span>.</div> <div class="article_info">L Mark, OB Spiller, M Okroj, S Chanas, JA Aitken, SW Wong, B Damania, AM Blom and DJ Blackbourn. <span class="article_issue_date"></span>. <span class="article_title">Molecular characterization of the Rhesus Rhadinovirus (RRV) ORF4 gene and the complement control protein (RCP) it encodes.</span> (submitted)</div>

PY - 2006

Y1 - 2006

N2 - Viruses have developed several strategies to evade the complement system, a part of the immune system. The fourth open reading frame of the human virus Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a type 1 transmembrane protein. This contains four complement control protein domains (CCP domains), and is thus structurally similar to human complement inhibitors. We named it KSHV complement control protein (KCP). Using recombinant KCP, we showed that it can inhibit the complement system by disrupting a key enzymatic complex, the C3 convertase. We predicted the 3D structure of KCP by homology modeling. Functionally important sites in KCP were mapped using mutants where either one to three amino acids, or entire CCP domains, were altered. We also found that a patch of positively charged amino acids in CCP 1-2 of KCP can bind to heparin and weakly to heparin like proteoglycans at the surface of cells, which seems to aid the virus in the attachment step prior to cell entry. In order to study the biological role of complement inhibition for this type of virus, we want to use the KSHV homologue rhesus rhadinovirus (RRV), since this virus is both technically easier to work with than KCP and also permits in vivo studies, by naturally infecting Rhesus Macaques. RRV complement control protein (RCP) is the KCP homologue in RRV. We have characterized its gene, confirmed that RCP is expressed and present on the surface of virus and performed experiments indicating that it is a functional complement inhibitor.

AB - Viruses have developed several strategies to evade the complement system, a part of the immune system. The fourth open reading frame of the human virus Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a type 1 transmembrane protein. This contains four complement control protein domains (CCP domains), and is thus structurally similar to human complement inhibitors. We named it KSHV complement control protein (KCP). Using recombinant KCP, we showed that it can inhibit the complement system by disrupting a key enzymatic complex, the C3 convertase. We predicted the 3D structure of KCP by homology modeling. Functionally important sites in KCP were mapped using mutants where either one to three amino acids, or entire CCP domains, were altered. We also found that a patch of positively charged amino acids in CCP 1-2 of KCP can bind to heparin and weakly to heparin like proteoglycans at the surface of cells, which seems to aid the virus in the attachment step prior to cell entry. In order to study the biological role of complement inhibition for this type of virus, we want to use the KSHV homologue rhesus rhadinovirus (RRV), since this virus is both technically easier to work with than KCP and also permits in vivo studies, by naturally infecting Rhesus Macaques. RRV complement control protein (RCP) is the KCP homologue in RRV. We have characterized its gene, confirmed that RCP is expressed and present on the surface of virus and performed experiments indicating that it is a functional complement inhibitor.

KW - transplantation

KW - serologi

KW - serology

KW - Kaposi's sarcoma-associated herpesvirus

KW - Immunologi

KW - Immunology

KW - homology modeling

KW - complement inhibitor

M3 - Doctoral Thesis (compilation)

SN - 91-85559-62-8

T3 - 2006:140

PB - Department of Laboratory Medicine, Lund University

ER -

Structural and functional studies of novel viral complement inhibitors

Abstract

Bibliographical note

Subject classification (UKÄ)

Free keywords

Fingerprint

Cite this