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Extraordinary Diversity of Immune Response Proteins among Sea Urchins: Nickel-Isolated Sp185/333 Proteins Show Broad Variations in Size and Charge.

Sherman LS, Schrankel CS, Brown KJ, Smith LC - PLoS ONE (2015)

Bottom Line: Effective protection against pathogens requires the host to produce a wide range of immune effector proteins.The Ni-Sp185/333 protein repertoire is variable among animals, and shows a variety of changes among individual sea urchins in response to immune challenges with both the same and different species of bacteria.The extraordinary diversity of the Ni-Sp185/333 proteins may provide significant anti-pathogen capabilities for sea urchins that survive solely on innate immunity.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, George Washington University, Washington DC, United States of America.

ABSTRACT
Effective protection against pathogens requires the host to produce a wide range of immune effector proteins. The Sp185/333 gene family, which is expressed by the California purple sea urchin Strongylocentrotus purpuratus in response to bacterial infection, encodes a highly diverse repertoire of anti-pathogen proteins. A subset of these proteins can be isolated by affinity to metal ions based on multiple histidines, resulting in one to four bands of unique molecular weight on standard Western blots, which vary depending on the individual sea urchin. Two dimensional gel electrophoresis (2DE) of nickel-isolated protein samples followed by Western blot was employed to detect nickel-isolated Sp185/333 (Ni-Sp185/333) proteins and to evaluate protein diversity in animals before and after immune challenge with marine bacteria. Ni-Sp185/333 proteins of the same molecular weight on standard Western blots appear as a broad complex of variants that differ in pI on 2DE Western blots. The Ni-Sp185/333 protein repertoire is variable among animals, and shows a variety of changes among individual sea urchins in response to immune challenges with both the same and different species of bacteria. The extraordinary diversity of the Ni-Sp185/333 proteins may provide significant anti-pathogen capabilities for sea urchins that survive solely on innate immunity.

No MeSH data available.


Related in: MedlinePlus

Major changes in the Ni-Sp185/333 protein repertoire may not occur until after the second challenge with V. diazotrophicus.(A) Sea urchin 106 has a limited array of Ni-Sp185/333 proteins prior to immune challenge, with a few high MW proteins of basic pI (arrow). (B) After the first challenge with V. diazotrophicus, the Ni-Sp185/333 proteins appear as more acidic (arrows) plus proteins of ~60 kDa appear (arrow head). (C) After the second challenge with V. diazotrophicus, a significant change in the repertoire of Ni-Sp185/333 proteins shows a shift to more basic pI for both the large MW proteins (arrows), and the ~60 kDa proteins (arrowhead). These images were cropped on the bottom because they do not show spots of less than 50 kDa.
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pone.0138892.g008: Major changes in the Ni-Sp185/333 protein repertoire may not occur until after the second challenge with V. diazotrophicus.(A) Sea urchin 106 has a limited array of Ni-Sp185/333 proteins prior to immune challenge, with a few high MW proteins of basic pI (arrow). (B) After the first challenge with V. diazotrophicus, the Ni-Sp185/333 proteins appear as more acidic (arrows) plus proteins of ~60 kDa appear (arrow head). (C) After the second challenge with V. diazotrophicus, a significant change in the repertoire of Ni-Sp185/333 proteins shows a shift to more basic pI for both the large MW proteins (arrows), and the ~60 kDa proteins (arrowhead). These images were cropped on the bottom because they do not show spots of less than 50 kDa.

Mentions: The third category of outcomes presented arrays of Ni-Sp185/333 proteins that shifted MW and/or pI after challenge. Several examples in this category were observed including sea urchin 102, which yielded similar numbers of Ni-Sp185/333 MW/pI variants of the same MW, but which shifted after challenge to more basic pI with an increased number of variants (box; Fig 4B). Sea urchin 103 showed a similar shift towards more basic isoforms for proteins of 150 to 250 kDa (white boxes, white arrows; Fig 5A and 5B), but also showed changes in protein abundance (black arrows; Fig 5A and 5B). Sea urchin 106 had unique MW/pI Sp185/333 spots that were present both before and after a single challenge with V. diazotrophicus that expanded towards acidic pI (arrows; Fig 8A and 8B) in addition to the appearance of ~55 kDa proteins and a slightly basic spot (arrow head; Fig 8B). Sea urchin 107 showed a decrease in the repertoire of Ni-Sp185/333 proteins after the first challenge with V. diazotrophicus compared to pre-challenge, with shifts towards more basic proteins in the major trains (white arrows; Fig 9A and 9B) and the appearance of more acidic trains after challenge (black arrowheads, Fig 9B). In general, an increase in Ni-Sp185/333 proteins following an initial challenge with V. diazotrophicus was consistent with reported changes in gene expression [7] in addition to a decrease in edited Sp185/333 cDNA sequences encoding truncated proteins that was noted in 8 of 11 sea urchins following immune challenge or injury (Fig 2). Overall, a wide range of Ni-Sp185/333 protein repertoires were observed among sea urchins, which illustrated the diversity of the system among animals and the diversity of responses to the same bacterial challenge.


Extraordinary Diversity of Immune Response Proteins among Sea Urchins: Nickel-Isolated Sp185/333 Proteins Show Broad Variations in Size and Charge.

Sherman LS, Schrankel CS, Brown KJ, Smith LC - PLoS ONE (2015)

Major changes in the Ni-Sp185/333 protein repertoire may not occur until after the second challenge with V. diazotrophicus.(A) Sea urchin 106 has a limited array of Ni-Sp185/333 proteins prior to immune challenge, with a few high MW proteins of basic pI (arrow). (B) After the first challenge with V. diazotrophicus, the Ni-Sp185/333 proteins appear as more acidic (arrows) plus proteins of ~60 kDa appear (arrow head). (C) After the second challenge with V. diazotrophicus, a significant change in the repertoire of Ni-Sp185/333 proteins shows a shift to more basic pI for both the large MW proteins (arrows), and the ~60 kDa proteins (arrowhead). These images were cropped on the bottom because they do not show spots of less than 50 kDa.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4583492&req=5

pone.0138892.g008: Major changes in the Ni-Sp185/333 protein repertoire may not occur until after the second challenge with V. diazotrophicus.(A) Sea urchin 106 has a limited array of Ni-Sp185/333 proteins prior to immune challenge, with a few high MW proteins of basic pI (arrow). (B) After the first challenge with V. diazotrophicus, the Ni-Sp185/333 proteins appear as more acidic (arrows) plus proteins of ~60 kDa appear (arrow head). (C) After the second challenge with V. diazotrophicus, a significant change in the repertoire of Ni-Sp185/333 proteins shows a shift to more basic pI for both the large MW proteins (arrows), and the ~60 kDa proteins (arrowhead). These images were cropped on the bottom because they do not show spots of less than 50 kDa.
Mentions: The third category of outcomes presented arrays of Ni-Sp185/333 proteins that shifted MW and/or pI after challenge. Several examples in this category were observed including sea urchin 102, which yielded similar numbers of Ni-Sp185/333 MW/pI variants of the same MW, but which shifted after challenge to more basic pI with an increased number of variants (box; Fig 4B). Sea urchin 103 showed a similar shift towards more basic isoforms for proteins of 150 to 250 kDa (white boxes, white arrows; Fig 5A and 5B), but also showed changes in protein abundance (black arrows; Fig 5A and 5B). Sea urchin 106 had unique MW/pI Sp185/333 spots that were present both before and after a single challenge with V. diazotrophicus that expanded towards acidic pI (arrows; Fig 8A and 8B) in addition to the appearance of ~55 kDa proteins and a slightly basic spot (arrow head; Fig 8B). Sea urchin 107 showed a decrease in the repertoire of Ni-Sp185/333 proteins after the first challenge with V. diazotrophicus compared to pre-challenge, with shifts towards more basic proteins in the major trains (white arrows; Fig 9A and 9B) and the appearance of more acidic trains after challenge (black arrowheads, Fig 9B). In general, an increase in Ni-Sp185/333 proteins following an initial challenge with V. diazotrophicus was consistent with reported changes in gene expression [7] in addition to a decrease in edited Sp185/333 cDNA sequences encoding truncated proteins that was noted in 8 of 11 sea urchins following immune challenge or injury (Fig 2). Overall, a wide range of Ni-Sp185/333 protein repertoires were observed among sea urchins, which illustrated the diversity of the system among animals and the diversity of responses to the same bacterial challenge.

Bottom Line: Effective protection against pathogens requires the host to produce a wide range of immune effector proteins.The Ni-Sp185/333 protein repertoire is variable among animals, and shows a variety of changes among individual sea urchins in response to immune challenges with both the same and different species of bacteria.The extraordinary diversity of the Ni-Sp185/333 proteins may provide significant anti-pathogen capabilities for sea urchins that survive solely on innate immunity.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, George Washington University, Washington DC, United States of America.

ABSTRACT
Effective protection against pathogens requires the host to produce a wide range of immune effector proteins. The Sp185/333 gene family, which is expressed by the California purple sea urchin Strongylocentrotus purpuratus in response to bacterial infection, encodes a highly diverse repertoire of anti-pathogen proteins. A subset of these proteins can be isolated by affinity to metal ions based on multiple histidines, resulting in one to four bands of unique molecular weight on standard Western blots, which vary depending on the individual sea urchin. Two dimensional gel electrophoresis (2DE) of nickel-isolated protein samples followed by Western blot was employed to detect nickel-isolated Sp185/333 (Ni-Sp185/333) proteins and to evaluate protein diversity in animals before and after immune challenge with marine bacteria. Ni-Sp185/333 proteins of the same molecular weight on standard Western blots appear as a broad complex of variants that differ in pI on 2DE Western blots. The Ni-Sp185/333 protein repertoire is variable among animals, and shows a variety of changes among individual sea urchins in response to immune challenges with both the same and different species of bacteria. The extraordinary diversity of the Ni-Sp185/333 proteins may provide significant anti-pathogen capabilities for sea urchins that survive solely on innate immunity.

No MeSH data available.


Related in: MedlinePlus