Influence of short peptides with aromatic amino acid residues on aggregation properties of serum amyloid A and its fragments

Mast cells are known to accumulate in various inflammatory processes, some of which are known to be associated with increased local and systemic levels of acute-phase reactants such as serum amyloid A (SAA) or with amyloid deposition. The mechanism(s) by which mast cells are recruited to these sites, however, has not been fully elucidated. It has recently been shown that SAA interacts with extracellular matrix (ECM) components and thereby acts as a chemoattractant and regulator of immune cell migration. On the basis of these observations, we examined the effect of SAA on mast cell adhesion to ECM, an essential step in cellular transmigration. We could first demonstrate strong specific binding of recombinant human SAA (rSAA) to murine mast cells using flow cytometry. Moreover, radiolabeled rSAA was found to bind, in a saturable manner, to mast cells, reaching a binding affinity of 10(-8) M. When immobilized by preincubation with ECM, SAA or its proteolytically degraded amyloid A fragment (amino acid residues 2-82), which contains RGD-related adhesion motif but not the COOH-terminal portion of SAA (amino acid residues 77-104), induced the adhesion of resting mast cells to ECM or laminin. SAA and AA, in soluble or immobilized forms, did not activate mast cells to release mediators. Mast cell adhesion to the immobilized ECM-SAA complex appeared to occur through an integrin recognition, inasmuch as adhesion was calcium dependent and could be blocked by an RGD-containing peptide or by anti-CD29 monoclonal antibody. Genistein also inhibited adhesion, indicating that tyrosine kinase activity was involved. These data suggest that SAA bound to ECM may serve as an important inducer of mast cell adhesion, thus regulating mast cell recruitment and accumulation at these sites, which in turn could potentiate further pathology.

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Aggregation of Mouse Serum Amyloid A Protein Was Promoted by Amyloid-Enhancing Factors with the More Genetically Homologous Serum Amyloid A

International Journal of Molecular Sciences ◽

10.3390/ijms22031036 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1036

Author(s):

Xuguang Lin ◽

Kenichi Watanabe ◽

Masahiro Kuragano ◽

Kiyotaka Tokuraku

Keyword(s):

Amino Acid ◽

Serum Amyloid A ◽

Animal Species ◽

Amino Acid Sequences ◽

Serum Amyloid ◽

Mouse Serum ◽

Aa Amyloidosis ◽

Basic Residue ◽

Amyloid A ◽

Serum Amyloid A Protein

Amyloid A (AA) amyloidosis is a condition in which amyloid fibrils characterized by a linear morphology and a cross-β structure accumulate and are deposited extracellularly in organs, resulting in chronic inflammatory diseases and infections. The incidence of AA amyloidosis is high in humans and several animal species. Serum amyloid A (SAA) is one of the most important precursor amyloid proteins and plays a vital step in AA amyloidosis. Amyloid enhancing factor (AEF) serves as a seed for fibril formation and shortens the onset of AA amyloidosis sharply. In this study, we examined whether AEFs extracted and purified from five animal species (camel, cat, cattle, goat, and mouse) could promote mouse SAA (mSAA) protein aggregation in vitro using quantum-dot (QD) nanoprobes to visualize the aggregation. The results showed that AEFs shortened and promoted mSAA aggregation. In addition, mouse and cat AEFs showed higher mSAA aggregation-promoting activity than the camel, cattle, and goat AEFs. Interestingly, homology analysis of SAA in these five animal species revealed a more similar amino acid sequence homology between mouse and cat than between other animal species. Furthermore, a detailed comparison of amino acid sequences suggested that it was important to mSAA aggregation-promoting activity that the 48th amino acid was a basic residue (Lys) and the 125th amino acid was an acidic residue (Asp or Glu). These data imply that AA amyloidosis exhibits higher transmission activity among animals carrying genetically homologous SAA gene, and may provide a new understanding of the pathogenesis of amyloidosis.

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Fibrous Aggregates of Short Peptides Containing Two Distinct Aromatic Amino Acid Residues

Chemistry & Biodiversity ◽

10.1002/cbdv.201900339 ◽

2019 ◽

Vol 16 (11) ◽

Cited By ~ 2

Author(s):

Wojciech Lipinski ◽

Joanna Wasko ◽

Malgorzata Walczak ◽

Justyna Fraczyk ◽

Zbigniew J. Kaminski ◽

...

Keyword(s):

Amino Acid ◽

Aromatic Amino Acid ◽

Short Peptides ◽

Amino Acid Residues

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Search for Fibrous Aggregates Potentially Useful in Regenerative Medicine Formed under Physiological Conditions by Self-Assembling Short Peptides Containing Two Identical Aromatic Amino Acid Residues

Molecules ◽

10.3390/molecules23030568 ◽

2018 ◽

Vol 23 (3) ◽

pp. 568 ◽

Cited By ~ 3

Author(s):

Justyna Fraczyk ◽

Wojciech Lipinski ◽

Agata Chaberska ◽

Joanna Wasko ◽

Kamil Rozniakowski ◽

...

Keyword(s):

Amino Acid ◽

Regenerative Medicine ◽

Aromatic Amino Acid ◽

Short Peptides ◽

Amino Acid Residues ◽

Physiological Conditions ◽

Self Assembling

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The Revised Amino Acid Sequence of Serum Amyloid A (SAA) Protein in Mink

Scandinavian Journal of Immunology ◽

10.1111/j.1365-3083.1993.tb01686.x ◽

1993 ◽

Vol 37 (6) ◽

pp. 696-697 ◽

Cited By ~ 13

Author(s):

P. V. SYVERSEN ◽

K. SLETTEN ◽

G. MARHAUG ◽

G. HUSBY ◽

B. LIUM

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Serum Amyloid A ◽

Serum Amyloid ◽

Amyloid A

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Effect of amino acid variations in the central region of human serum amyloid A on the amyloidogenic properties

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2014.01.029 ◽

2014 ◽

Vol 444 (1) ◽

pp. 92-97 ◽

Cited By ~ 16

Author(s):

Hiroka Takase ◽

Masafumi Tanaka ◽

Sachiko Miyagawa ◽

Toshiyuki Yamada ◽

Takahiro Mukai

Keyword(s):

Amino Acid ◽

Human Serum ◽

Central Region ◽

Serum Amyloid A ◽

Serum Amyloid ◽

Amyloid A

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Can Short Peptides Be Inhibitors of Serum Amyloid a Protein Aggregation?

Proceedings ◽

10.3390/proceedings2019022038 ◽

2019 ◽

Vol 22 (1) ◽

pp. 38

Author(s):

Skibiszewska ◽

Jankowska

Keyword(s):

Protein Aggregation ◽

Serum Amyloid A ◽

Serum Amyloid ◽

Short Peptides ◽

Amyloid A ◽

Serum Amyloid A Protein ◽

Amyloidogenic Protein ◽

Protein Molecules ◽

Self Association

One of the approaches in the design of anti-amyloid drugs is to find compounds that are able to hamper self-association of amyloidogenic protein molecules. [...]

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Expression of recombinant human serum amyloid A in mammalian cells and demonstration of the region necessary for high-density lipoprotein binding and amyloid fibril formation by site-directed mutagenesis

Biochemical Journal ◽

10.1042/bj3181041 ◽

1996 ◽

Vol 318 (3) ◽

pp. 1041-1049 ◽

Cited By ~ 48

Author(s):

Himakshi PATEL ◽

Jo BRAMALL ◽

Helen WATERS ◽

Maria C. DE BEER ◽

Patricia WOO

Keyword(s):

Amino Acid ◽

Serum Amyloid A ◽

Amyloid Fibrils ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Site Directed Mutagenesis ◽

Amino Acid Residues ◽

Wild Type ◽

Amyloid A ◽

Amyloid Fibril Formation

Site-directed mutagenesis of the acute-phase human serum amyloid A (SAA1α) protein was used to evaluate the importance of the N-terminal amino acid residues, namely RSFFSFLGEAF. The full-length cDNA clone of SAA1α (pA1.mod.) was used to create two mutations, namely Gly-8 to Asp-8 and an 11 amino acid truncation between Arg-1 and Phe-11 respectively. Wild-type and mutant cDNAs were expressed in Chinese hamster ovary (CHO) cells under the control of the human cytomegalovirus promoter, which resulted in the secretion of the processed proteins into the culture media. Wild-type recombinant human SAA (rSAA) protein was shown to have pI values of 6.0 and 6.4, similar to the human SAA isoform SAA1α and SAA1α desArg found in acute-phase plasma. N-terminal sequencing of 56 residues confirmed its identity with human SAA1α. The total yield of wild-type rSAA measured by ELISA was between 3.5 and 30 mg/l. The two mutations resulted in reduced expression levels of the mutant SAA proteins (3–10 mg/l). Further measurements of rSAA concentration in lipid fractions of culture medium collected at a density of 1.21 g/ml (high-density lipoprotein; HDL) and 1.063–1.18 g/ml (very-low-density lipoprotein/low-density lipoprotein; VLDL/LDL) showed that 76% of the wild-type protein was found in the HDL fraction and the remaining 24% in the infranatant non-lipid fraction. In contrast the relative concentration of mutant rSAA in HDL and infranatant fractions was reversed. This is consistent with the previously proposed involvement of the 11 amino acid peptide in anchoring SAA protein on to HDL3 [Turnell, Sarra, Glover, Baum, Caspi, Baltz and Pepys (1986) Mol. Biol. Med.3, 387–407]. Wild-type rSAA protein was shown to form amyloid fibrils in vitro under acidic conditions as shown by electron microscopy, and stained positive with Congo Red and exhibited apple-green birefringence when viewed under polarized light. Under the same conditions mutSAA(G8D) and mutSAAΔ1–11 did not form amyloid fibrils. In conclusion, replacement of Gly-8 by Asp-8 or deletion of the first 11 amino acid residues at the N-terminus of rSAA diminishes its capacity to bind to HDL and decreases amyloid fibril formation.

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Heterogeneity of human serum amyloid A proteins.

Journal of Experimental Medicine ◽

10.1084/jem.152.3.641 ◽

1980 ◽

Vol 152 (3) ◽

pp. 641-656 ◽

Cited By ~ 100

Author(s):

L L Bausserman ◽

P N Herbert ◽

K P McAdam

Keyword(s):

Amino Acid ◽

Serum Amyloid A ◽

Polyacrylamide Gel ◽

Serum Amyloid ◽

High Density Lipoproteins ◽

Systemic Lupus Erythematosis ◽

Secondary Amyloidosis ◽

Single Chain ◽

Amino Acid Compositions ◽

Amyloid A

Serum amyloid A proteins (SAA), presumed precursors of the tissue amyloid A proteins (AA) characteristic of secondary amyloidosis, have been isolated from the plasma high-density lipoproteins (HDL) of normals after etiocholanolone-induced inflammation and from patients with Wegener's granulomatosis, systemic lupus erythematosis, juvenile rheumatoid arthritis, Waldenström's macroglobulinemia, and Goodpasture's syndrome. At least six polymorphic forms of SAA wer identified among the low molecular weight proteins of HDL, and these comprosed up to 27% of the total HDL protein. Gel and ion-exchange chromatography permitted isolation of the SAA polymorphs in homogeneous form. Their amino acid compositions were very similar, they were indistinguishable in cationic and sodium dodecyl sulfate-polyacrylamide gel electrophoresis systems, and each had the terminal sequency COOH-Tyr-Lys-Phe-. Charge heterogeneity in anionic-urea polyacrylamide gel electropherograms was unaffected by neuaminidase treatment, and none of the SAA protein bands stained with the periodate-Schiff reagent. The two major SAA polymorphs, designated SAA4 and SAA5 according to their order of elution from DEAE-cellulose, had different NH2-terminal sequences. Manual Edman degradation demonstrated NH2-arg-ser-phe-phe- for SAA4 and NH2-ser-phe-phe- for SAA5. This NH2-terminal heterogeneity corresponds to that most frequently reported for AA and suggests that microheterogeneity in SAA may underlie that already documented in AA. Sufficient quantitites of the other SAA polymorphs were not available for similar analyses, but the amino acid compositions do not indicate that NH2-terminal heterogeneity accounts for all of the observed polymorphism. Artifactual polymorphism also appears unlikely, and the heterogeneiy of SAA may reflect origin from more than one cell type with or without posttranslational modificaton. We calculate from quantitative COOH-terminal analyses that SAA is of 11,000-11,900 mol wt. Primary structure studies have shown AA t be a single chain protein of 76 residues, and SAA, therefore, appears to contain a peptide of 33 amino acids that is missing from AA.

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