Structure and biological properties of scavenger receptor MARCO
Author: Brännström, Annika
Date: 2002-09-27
Location: Samuelssonsalen, Scheelelaboratoriet, Scheeles väg 2, Karolinska Institutet
Time: 9.00
Department: Institutionen för medicinsk biokemi och biofysik (MBB) / Department of Medical Biochemistry and Biophysics
View/ Open:
thesis.pdf (950.5Kb)
Abstract
Macrophages are monocyte-derived cells that play an important role in the
innate immune response against invading pathogens. These cells express
several host defense receptors that can be divided into two classes;
those dependent on opsonizing components for recognition of pathogens,
and those that can recognize pathogens directly, pattern recognition
receptors (PRRs). Class A scavenger receptors are a family of PRRs
composed of three members: Scavenger Receptor A (SRA), MAcrophage Receptor
with COllagenous structure (MARCO), and a recently identified protein Scavenger
Receptor with C-type Lectin (SRCL). MARCO is a trimeric membrane protein
containing an Nterminal intracellular domain, a transmembrane domain, and
an extracellular portion composed of a short spacer domain, a triple-
helical collagenous domain, and a C-terminal cysteine-rich domain (SRCR).
In unstimulated mice, MARCO expression is restricted to the marginal zone
macrophages in spleen, macrophages in medullary cord in lymph nodes, and
peritoneal macrophages. Its expression can be induced in cultured
macrophages or in macrophages of various tissues after administering LPS,
BCG, Listeria monocytogenes or zymosan. These findings suggest a role in
host defense. Furthermore, cells transfected with MARCO cDNA avidly bind
both gram-negative and gram-positive bacteria, but not yeast. Preliminary
data shows that the bacteria-binding region is in the SRCR domain.
Interestingly, MARCO was found to be the major receptor on alveolar
macrophages for binding of unopsonized environmental particles such as
TiO2 and Fe203
To examine the regulatory mechanisms of MARCO, and its potential role in
disease, the structures of human and murine MARCO genes were determined.
Both genes have 17 exons, of which exons 4-15 encode the collagenous
domain. Two major transcription initiation sites were identified, one
starting at position +1, 27 bp downstream of a TATA box, and another at
position -64 downstream of an AT-rich region. Several potential binding
sites for transcription factors involved in host defense were identified
in the promoter region. The human and mouse genes were localized to
syntenic regions on chromosomes 2 and 1, respectively.
We could also show that MARCO most likely has a direct effect on the
phenotype of activated macrophages, since the expression of MARCO in
different cell lines induces dramatic cell shape changes. Typically these
changes include formation of large lamellipodia-like structures and long
dendritic processes. The morphological changes are accompanied by
disassembly of actin stress fibers and often also by complete loss of
focal adhesions. The MARCO- induced changes are dependent on cell
adhesion and are inhibited when the cells are plated on
fibronectin-coated surfaces. Similarly, a dominant-negative mutant of
Rac- I partially inhibits the morphogenic effects of MARCO in CHO cells.
Our data indicate that the proximal segment of the SRCR domain is
important for the morphoregulatory activity.
We used a large number of human and mouse MARCO variants to show that the
predominant bacteria- binding region of MARCO resides in the SRCR domain.
This result demonstrates the first function for a SRCR domain of the
scavenger receptor A family. In further analysis, we found that an
arginine-rich segment in the cysteine-rich domain is responsible for the
high-affinity binding. More precisely, the motif arginine-X-arginine was
found to be of importance in this regard. This was demonstrated by
comparing the bacteria-binding activity of two truncated constructs of
MARCO. One form, which contains an argininedoublet in its SRCR domain,
binds bacteria avidly, while the other, containing one single arginine
residue, exhibits insignificant binding.
To obtain a tool for ligand-binding studies, we produced the
extracellular part of MARCO as a recombinant protein, sMARCO. Several
studies indicated that sMARCO has a triple-helical structure. When imaged
in electron microscope after rotary shadowing, the protein appeared as
82.7 nm long rod-like molecules with small globes at both ends.
Furthermore, the molecules were often found to be associated with each
other. Binding studies with sMARCO, as well as with MARCO-expressing
cells, indicated that MARCO binds gram-negative strains expressing either
the smooth- or rough- form of I VS. Additional studies revealed that LPS
is a ligand of MARCO. Finally, we also produced the SRCR domain as a
recombinant protein, and compared the bacteria-binding properties of this
protein and sMARCO. These studies indicated that the SRCR domain has to
be in a trimeric form in order to bind bacteria efficiently.
List of papers:
I. Kangas M, Brannstrom A, Elomaa O, Matsuda Y, Eddy R, Shows TB, Tryggvason K (1998). "Structure and chromosomal localization of the human and murine genes for the macrophage MARCO receptor. " Genomics 58(1): 82-9
Pubmed
II. Pikkarainen T, Brannstrom A, Tryggvason K (1999). "Expression of macrophage MARCO receptor induces formation of dendritic plasma membrane processes. " J Biol Chem 274(16): 10975-82
Pubmed
III. Brannstrom A, Sankala M, Tryggvason K, Pikkarainen T (2002). "Arginine residues in domain V have a central role for bacteria-binding activity of macrophage scavenger receptor MARCO. " Biochem Biophys Res Commun 290(5): 1462-9
Pubmed
IV. Sankala M, Brannstrom A, Schulthess T, Bergmann U, Morgunova E, Engel J, Tryggvason K, Pikkarainen T (2002). "Characterization of Recombinant Soluble Macrophage Scavenger Receptor MARCO. " J Biol Chem 277(36): 33378-33385
Pubmed
I. Kangas M, Brannstrom A, Elomaa O, Matsuda Y, Eddy R, Shows TB, Tryggvason K (1998). "Structure and chromosomal localization of the human and murine genes for the macrophage MARCO receptor. " Genomics 58(1): 82-9
Pubmed
II. Pikkarainen T, Brannstrom A, Tryggvason K (1999). "Expression of macrophage MARCO receptor induces formation of dendritic plasma membrane processes. " J Biol Chem 274(16): 10975-82
Pubmed
III. Brannstrom A, Sankala M, Tryggvason K, Pikkarainen T (2002). "Arginine residues in domain V have a central role for bacteria-binding activity of macrophage scavenger receptor MARCO. " Biochem Biophys Res Commun 290(5): 1462-9
Pubmed
IV. Sankala M, Brannstrom A, Schulthess T, Bergmann U, Morgunova E, Engel J, Tryggvason K, Pikkarainen T (2002). "Characterization of Recombinant Soluble Macrophage Scavenger Receptor MARCO. " J Biol Chem 277(36): 33378-33385
Pubmed
Issue date: 2002-09-06
Rights:
Publication year: 2002
ISBN: 91-7349-294-9
Statistics
Total Visits
Views | |
---|---|
Structure ...(legacy) | 648 |
Structure ... | 148 |
Total Visits Per Month
October 2023 | November 2023 | December 2023 | January 2024 | February 2024 | March 2024 | April 2024 | |
---|---|---|---|---|---|---|---|
Structure ... | 2 | 1 | 1 | 7 | 0 | 1 | 1 |
File Visits
Views | |
---|---|
thesis.pdf(legacy) | 353 |
thesis.pdf | 49 |
thesis.pdf.txt(legacy) | 2 |
Top country views
Views | |
---|---|
United States | 374 |
China | 65 |
Germany | 59 |
Sweden | 57 |
South Korea | 15 |
Russia | 13 |
Canada | 11 |
Finland | 10 |
United Kingdom | 10 |
India | 9 |
Top cities views
Views | |
---|---|
Sunnyvale | 30 |
Romeo | 28 |
Beijing | 18 |
Kiez | 18 |
Shenzhen | 17 |
Seoul | 15 |
Hamilton | 7 |
London | 7 |
Ashburn | 6 |
Ballerup | 6 |