Functional and structural diversities of C-reactive proteins present in horseshoe crab hemolymph plasma

Daisuke Iwaki, Tsukasa Osaki, Yoshimitsu Mizunoe, Sun N. Wai, Sadaaki Iwanaga, Shun Ichiro Kawabata

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Limulin, a sialic-acid-binding and phosphorylethanolamine-binding hemagglutinin in the hemolymph plasma of the American horseshoe crab (Limulus polyphemus), is a hemolytic C-reactive protein [Armstrong, P.B., Swarnakar, S., Srimal, S., Misquith, S., Hahn, E.A., Aimes, R.T. and Quigley, J.P. (1996) J. Biol. Chem. 271, 14717-14721]. We have now identified three types of C-reactive protein in the plasma of the Japanese horseshoe crab (Tachypleus tridentatus), based on different affinities against fetuin- agarose and phosphorylethanolamine-agarose determined by quantitative precipitin assays using fetuin and an artificial phosphorylethanolamine- protein conjugate. Partial amino acid sequences of the isolated C-reactive proteins identified homologous proteins which were named Tachypleus tridentatus CRP-1 (tCRP-1), tCRP-2 and tCRP-3, each of which possibly constitute isoprotein mixtures. tCRP-2 and tCRP-3, but not tCRP-1, agglutinated mammalian erythrocytes tCRP-1, the most abundant C-reactive protein in the plasma, exhibited the highest affinity to the phosphorylethanolamine-protein conjugate but lacked both sialic-acid-binding and hemolytic activities, tCRP-2 bound to both fetuin-agarose and phosphorylethanolamine-agarose, and exhibited Ca2+-dependent hemolytic and sialic-acid-binding activities, suggestive of limulin-like properties. Furthermore, tCRP-2 exhibited a higher affinity to colominic acid, a bacterial polysialic acid. By contrast, tCRP-3 shows stronger hemolytic, sialic-acid-binding and hemagglutinating activities than tCRP-2. tCRP-3 has no affinity to phosphorylethanolamine-agarose, phosphorylethanolamine-protein conjugate and colominic acid. This suggests tCRP-3 is a novel hemolytic C- reactive protein lacking a common characteristic of phosphorylethanolamine- agarose binding affinity. Twenty-two clones of tCRPs with different deduced amino acid sequences were obtained by PCR using oligonucleotide primers based on the N-terminal and C-terminal sequences of tCRPs and with templates including genomic DNA and cDNA of hemocytes or hepatopancreas derived from one individual. The translation products of the tCRP clones possess high molecular diversity which falls into three related groups, consistent with classification based on their biological activities. Only tCRP-3 contained a unique hydrophobic nonapeptide sequence that appears in the transmembrane domain of a major histocompatibility complex class I heavy chain of rainbow trout, suggesting the importance of the hydrophobic patch to the hemolytic activity of tCRP-3. The structural and functional diversities of tCRPs provide a good model for studying the properties of innate immunity in invertebrates, which survive without the benefit of acquired immunity.

Original languageEnglish
Pages (from-to)314-326
Number of pages13
JournalEuropean Journal of Biochemistry
Volume264
Issue number2
DOIs
Publication statusPublished - Sep 1 1999

Fingerprint

Horseshoe Crabs
Hemolymph
C-Reactive Protein
Sepharose
Plasmas
N-Acetylneuraminic Acid
Fetuins
Amino Acid Sequence
Proteins
Clone Cells
Hepatopancreas
Amino Acids
Hemocytes
Precipitins
DNA Primers
phosphorylethanolamine
Oncorhynchus mykiss
Hemagglutinins
Adaptive Immunity
Invertebrates

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Functional and structural diversities of C-reactive proteins present in horseshoe crab hemolymph plasma. / Iwaki, Daisuke; Osaki, Tsukasa; Mizunoe, Yoshimitsu; Wai, Sun N.; Iwanaga, Sadaaki; Kawabata, Shun Ichiro.

In: European Journal of Biochemistry, Vol. 264, No. 2, 01.09.1999, p. 314-326.

Research output: Contribution to journalArticle

Iwaki, Daisuke ; Osaki, Tsukasa ; Mizunoe, Yoshimitsu ; Wai, Sun N. ; Iwanaga, Sadaaki ; Kawabata, Shun Ichiro. / Functional and structural diversities of C-reactive proteins present in horseshoe crab hemolymph plasma. In: European Journal of Biochemistry. 1999 ; Vol. 264, No. 2. pp. 314-326.
@article{2aeb0e5edcb24effbd187782ef71d1cb,
title = "Functional and structural diversities of C-reactive proteins present in horseshoe crab hemolymph plasma",
abstract = "Limulin, a sialic-acid-binding and phosphorylethanolamine-binding hemagglutinin in the hemolymph plasma of the American horseshoe crab (Limulus polyphemus), is a hemolytic C-reactive protein [Armstrong, P.B., Swarnakar, S., Srimal, S., Misquith, S., Hahn, E.A., Aimes, R.T. and Quigley, J.P. (1996) J. Biol. Chem. 271, 14717-14721]. We have now identified three types of C-reactive protein in the plasma of the Japanese horseshoe crab (Tachypleus tridentatus), based on different affinities against fetuin- agarose and phosphorylethanolamine-agarose determined by quantitative precipitin assays using fetuin and an artificial phosphorylethanolamine- protein conjugate. Partial amino acid sequences of the isolated C-reactive proteins identified homologous proteins which were named Tachypleus tridentatus CRP-1 (tCRP-1), tCRP-2 and tCRP-3, each of which possibly constitute isoprotein mixtures. tCRP-2 and tCRP-3, but not tCRP-1, agglutinated mammalian erythrocytes tCRP-1, the most abundant C-reactive protein in the plasma, exhibited the highest affinity to the phosphorylethanolamine-protein conjugate but lacked both sialic-acid-binding and hemolytic activities, tCRP-2 bound to both fetuin-agarose and phosphorylethanolamine-agarose, and exhibited Ca2+-dependent hemolytic and sialic-acid-binding activities, suggestive of limulin-like properties. Furthermore, tCRP-2 exhibited a higher affinity to colominic acid, a bacterial polysialic acid. By contrast, tCRP-3 shows stronger hemolytic, sialic-acid-binding and hemagglutinating activities than tCRP-2. tCRP-3 has no affinity to phosphorylethanolamine-agarose, phosphorylethanolamine-protein conjugate and colominic acid. This suggests tCRP-3 is a novel hemolytic C- reactive protein lacking a common characteristic of phosphorylethanolamine- agarose binding affinity. Twenty-two clones of tCRPs with different deduced amino acid sequences were obtained by PCR using oligonucleotide primers based on the N-terminal and C-terminal sequences of tCRPs and with templates including genomic DNA and cDNA of hemocytes or hepatopancreas derived from one individual. The translation products of the tCRP clones possess high molecular diversity which falls into three related groups, consistent with classification based on their biological activities. Only tCRP-3 contained a unique hydrophobic nonapeptide sequence that appears in the transmembrane domain of a major histocompatibility complex class I heavy chain of rainbow trout, suggesting the importance of the hydrophobic patch to the hemolytic activity of tCRP-3. The structural and functional diversities of tCRPs provide a good model for studying the properties of innate immunity in invertebrates, which survive without the benefit of acquired immunity.",
author = "Daisuke Iwaki and Tsukasa Osaki and Yoshimitsu Mizunoe and Wai, {Sun N.} and Sadaaki Iwanaga and Kawabata, {Shun Ichiro}",
year = "1999",
month = "9",
day = "1",
doi = "10.1046/j.1432-1327.1999.00588.x",
language = "English",
volume = "264",
pages = "314--326",
journal = "FEBS Journal",
issn = "1742-464X",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - Functional and structural diversities of C-reactive proteins present in horseshoe crab hemolymph plasma

AU - Iwaki, Daisuke

AU - Osaki, Tsukasa

AU - Mizunoe, Yoshimitsu

AU - Wai, Sun N.

AU - Iwanaga, Sadaaki

AU - Kawabata, Shun Ichiro

PY - 1999/9/1

Y1 - 1999/9/1

N2 - Limulin, a sialic-acid-binding and phosphorylethanolamine-binding hemagglutinin in the hemolymph plasma of the American horseshoe crab (Limulus polyphemus), is a hemolytic C-reactive protein [Armstrong, P.B., Swarnakar, S., Srimal, S., Misquith, S., Hahn, E.A., Aimes, R.T. and Quigley, J.P. (1996) J. Biol. Chem. 271, 14717-14721]. We have now identified three types of C-reactive protein in the plasma of the Japanese horseshoe crab (Tachypleus tridentatus), based on different affinities against fetuin- agarose and phosphorylethanolamine-agarose determined by quantitative precipitin assays using fetuin and an artificial phosphorylethanolamine- protein conjugate. Partial amino acid sequences of the isolated C-reactive proteins identified homologous proteins which were named Tachypleus tridentatus CRP-1 (tCRP-1), tCRP-2 and tCRP-3, each of which possibly constitute isoprotein mixtures. tCRP-2 and tCRP-3, but not tCRP-1, agglutinated mammalian erythrocytes tCRP-1, the most abundant C-reactive protein in the plasma, exhibited the highest affinity to the phosphorylethanolamine-protein conjugate but lacked both sialic-acid-binding and hemolytic activities, tCRP-2 bound to both fetuin-agarose and phosphorylethanolamine-agarose, and exhibited Ca2+-dependent hemolytic and sialic-acid-binding activities, suggestive of limulin-like properties. Furthermore, tCRP-2 exhibited a higher affinity to colominic acid, a bacterial polysialic acid. By contrast, tCRP-3 shows stronger hemolytic, sialic-acid-binding and hemagglutinating activities than tCRP-2. tCRP-3 has no affinity to phosphorylethanolamine-agarose, phosphorylethanolamine-protein conjugate and colominic acid. This suggests tCRP-3 is a novel hemolytic C- reactive protein lacking a common characteristic of phosphorylethanolamine- agarose binding affinity. Twenty-two clones of tCRPs with different deduced amino acid sequences were obtained by PCR using oligonucleotide primers based on the N-terminal and C-terminal sequences of tCRPs and with templates including genomic DNA and cDNA of hemocytes or hepatopancreas derived from one individual. The translation products of the tCRP clones possess high molecular diversity which falls into three related groups, consistent with classification based on their biological activities. Only tCRP-3 contained a unique hydrophobic nonapeptide sequence that appears in the transmembrane domain of a major histocompatibility complex class I heavy chain of rainbow trout, suggesting the importance of the hydrophobic patch to the hemolytic activity of tCRP-3. The structural and functional diversities of tCRPs provide a good model for studying the properties of innate immunity in invertebrates, which survive without the benefit of acquired immunity.

AB - Limulin, a sialic-acid-binding and phosphorylethanolamine-binding hemagglutinin in the hemolymph plasma of the American horseshoe crab (Limulus polyphemus), is a hemolytic C-reactive protein [Armstrong, P.B., Swarnakar, S., Srimal, S., Misquith, S., Hahn, E.A., Aimes, R.T. and Quigley, J.P. (1996) J. Biol. Chem. 271, 14717-14721]. We have now identified three types of C-reactive protein in the plasma of the Japanese horseshoe crab (Tachypleus tridentatus), based on different affinities against fetuin- agarose and phosphorylethanolamine-agarose determined by quantitative precipitin assays using fetuin and an artificial phosphorylethanolamine- protein conjugate. Partial amino acid sequences of the isolated C-reactive proteins identified homologous proteins which were named Tachypleus tridentatus CRP-1 (tCRP-1), tCRP-2 and tCRP-3, each of which possibly constitute isoprotein mixtures. tCRP-2 and tCRP-3, but not tCRP-1, agglutinated mammalian erythrocytes tCRP-1, the most abundant C-reactive protein in the plasma, exhibited the highest affinity to the phosphorylethanolamine-protein conjugate but lacked both sialic-acid-binding and hemolytic activities, tCRP-2 bound to both fetuin-agarose and phosphorylethanolamine-agarose, and exhibited Ca2+-dependent hemolytic and sialic-acid-binding activities, suggestive of limulin-like properties. Furthermore, tCRP-2 exhibited a higher affinity to colominic acid, a bacterial polysialic acid. By contrast, tCRP-3 shows stronger hemolytic, sialic-acid-binding and hemagglutinating activities than tCRP-2. tCRP-3 has no affinity to phosphorylethanolamine-agarose, phosphorylethanolamine-protein conjugate and colominic acid. This suggests tCRP-3 is a novel hemolytic C- reactive protein lacking a common characteristic of phosphorylethanolamine- agarose binding affinity. Twenty-two clones of tCRPs with different deduced amino acid sequences were obtained by PCR using oligonucleotide primers based on the N-terminal and C-terminal sequences of tCRPs and with templates including genomic DNA and cDNA of hemocytes or hepatopancreas derived from one individual. The translation products of the tCRP clones possess high molecular diversity which falls into three related groups, consistent with classification based on their biological activities. Only tCRP-3 contained a unique hydrophobic nonapeptide sequence that appears in the transmembrane domain of a major histocompatibility complex class I heavy chain of rainbow trout, suggesting the importance of the hydrophobic patch to the hemolytic activity of tCRP-3. The structural and functional diversities of tCRPs provide a good model for studying the properties of innate immunity in invertebrates, which survive without the benefit of acquired immunity.

UR - http://www.scopus.com/inward/record.url?scp=0033199575&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033199575&partnerID=8YFLogxK

U2 - 10.1046/j.1432-1327.1999.00588.x

DO - 10.1046/j.1432-1327.1999.00588.x

M3 - Article

C2 - 10491075

AN - SCOPUS:0033199575

VL - 264

SP - 314

EP - 326

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 2

ER -