A proteinaceous organic matrix regulates carbonate mineral production in the marine teleost intestine.
Marine teleost fish produce CaCO3in their intestine as part of their osmoregulatory strategy. This precipitation is critical for rehydration and survival of the largest vertebrate group on earth, yet the molecular mechanisms that regulate this reaction are unknown. Here, we isolate and characterize an organic matrix associated with the intestinal precipitates produced by Gulf toadfish (Opsanus beta). Toadfish precipitates were purified using two different methods, and the associated organic matrix was extracted. Greater than 150 proteins were identified in the isolated matrix by mass spectrometry and subsequent database searching using an O. beta transcriptomic sequence library produced here. Many of the identified proteins were enriched in the matrix compared to the intestinal fluid, and three showed no substantial homology to any previously characterized protein in the NCBI database. To test the functionality of the isolated matrix, a micro-modified in vitro calcification assay was designed, which revealed that low concentrations of isolated matrix substantially promoted CaCO3production, where high concentrations showed an inhibitory effect. High concentrations of matrix also decreased the incorporation of magnesium into the forming mineral, potentially providing an explanation for the variability in magnesium content observed in precipitates produced by different fish species.
Marine Biology and Ecology, Rosenstiel School of Marine and Atmospheric Science, University of Miami, Miami, FL, 33149, USA.
Kevin L
KL
Schauer
Department of Biology, Brandon University, Brandon, MB, R7A 6A9, Canada.|Canadian Institute for Advanced Research, Department of Biology, University of Ottawa, Ottawa, ON, K1N 6N5, Canada.
Christophe M R
CM
LeMoine
Canadian Institute for Advanced Research, Department of Biology, University of Ottawa, Ottawa, ON, K1N 6N5, Canada.
Adrian
A
Pelin
Canadian Institute for Advanced Research, Department of Biology, University of Ottawa, Ottawa, ON, K1N 6N5, Canada.
Nicolas
N
Corradi
McDonnell Genome Institute, Washington University School of Medicine, St Louis, Missouri 63108, USA.
Wesley C
WC
Warren
Marine Biology and Ecology, Rosenstiel School of Marine and Atmospheric Science, University of Miami, Miami, FL, 33149, USA.
Martin
M
Grosell
M Danielle
MD
McDonald
D000818:Animals; D024541:Batrachoidiformes / Q000235:genetics / Q000378:metabolism*; D002119:Calcium Carbonate / Q000378:metabolism*; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism*; D007408:Intestinal Absorption; D007413:Intestinal Mucosa / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D013058:Mass Spectrometry
D029941:Fish Proteins; D002119:Calcium Carbonate; D008274:Magnesium
Am J Physiol. 1999 Dec;277(6):G1108-37
10600809
J Biol Chem. 2000 Jan 14;275(2):1057-64
10625646
Bioinformatics. 2001 Mar;17(3):282-3
11294794
Science. 1975 Dec 5;190(4218):987-9
1188379
Biochim Biophys Acta. 2002 Nov 13;1566(1-2):182-93
12421549
Curr Rheumatol Rep. 2003 Jun;5(3):222-6
12744815
Comp Biochem Physiol A Mol Integr Physiol. 2003 Apr;134(4):763-76
12814785
Biochim Biophys Acta. 2003 Dec 30;1618(2):163-74
14729153
Bioinformatics. 2005 Sep 15;21(18):3674-6
16081474
Biophys J. 2006 Oct 1;91(7):2601-8
16861275
Proc Natl Acad Sci U S A. 2006 Dec 19;103(51):19237-42
17158220
Biophys J. 2007 Aug 15;93(4):1246-54
17496038
Science. 1981 Jun 19;212(4501):1397-8
17746262
Biotechnol Genet Eng Rev. 2006;22:1-19
18476323
Science. 2008 Oct 31;322(5902):724-7
18974348
Chem Rev. 2008 Nov;108(11):4551-627
19006398
Science. 2009 Jan 16;323(5912):359-62
19150840
Am J Physiol Regul Integr Comp Physiol. 2010 Jul;299(1):R150-8
20410471
Am J Physiol Regul Integr Comp Physiol. 2011 Apr;300(4):R895-909
21228343
Acta Physiol (Oxf). 2011 Jul;202(3):421-34
21362153
Proc Natl Acad Sci U S A. 2011 Mar 8;108(10):3865-9
21368155
Bioinformatics. 2012 Apr 15;28(8):1086-92
22368243
BMC Bioinformatics. 2012 May 24;13:111
22624656
J Exp Biol. 2012 Nov 1;215(Pt 21):3836-44
22855618
Bioinformatics. 2012 Sep 15;28(18):i458-i465
22962467
Connect Tissue Res. 2012;53(6):438-45
22992051
Proc Natl Acad Sci U S A. 2012 Dec 18;109(51):20986-91
23213212
Cell Tissue Res. 2013 Jul;353(1):123-37
23644767
Am J Physiol Renal Physiol. 2013 Sep 1;305(5):F628-40
23804448
Connect Tissue Res. 2014 Jan-Feb;55(1):34-40
24437603
J Exp Biol. 2014 May 1;217(Pt 9):1555-62
24501133
Nucleic Acids Res. 2014 Jul;42(Web Server issue):W337-43
24799431
Proteome Sci. 2014 May 18;12:28
25018669
Am J Physiol Regul Integr Comp Physiol. 2014 Nov 1;307(9):R1167-79
25100079
Proc Natl Acad Sci U S A. 2014 Sep 2;111(35):12728-33
25139990
Nucleic Acids Res. 2015 Jan;43(Database issue):D222-6
25414356
Nucleic Acids Res. 2016 Jan 4;44(D1):D447-56
26527722
Proc Biol Sci. 2016 Apr 27;283(1829):
27122561
Proteomes. 2015 Aug 18;3(3):236-248
28248269
Biol Bull. 1990 Oct;179(2):191-200
29314985
Anal Biochem. 1985 Oct;150(1):76-85
3843705
J Exp Biol. 1974 Apr;60(2):535-46
4832995
Eur J Biochem. 1982 Feb;122(1):111-7
7060561
Proc Natl Acad Sci U S A. 1997 Aug 5;94(16):8411-6
9237990
J Exp Biol. 1996;199(Pt 10):2331-43
9320250