Review History


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Summary

  • The initial submission of this article was received on December 18th, 2017 and was peer-reviewed by 2 reviewers and the Academic Editor.
  • The Academic Editor made their initial decision on January 11th, 2018.
  • The first revision was submitted on February 20th, 2018 and was reviewed by 2 reviewers and the Academic Editor.
  • The article was Accepted by the Academic Editor on February 25th, 2018.

Version 0.2 (accepted)

· · Academic Editor

Accept

Due the potential use of the information provided in the field of molecular parasitology, the publication of this solid manuscript is recommended.

·

Basic reporting

I have read the revised manuscript and the authors have addressed my comments satisfactorily. I now recommend the manuscript for publication.

Experimental design

I have read the revised manuscript and the authors have addressed my comments satisfactorily. I now recommend the manuscript for publication.

Validity of the findings

I have read the revised manuscript and the authors have addressed my comments satisfactorily. I now recommend the manuscript for publication.

Comments for the author

I have read the revised manuscript and the authors have addressed my comments satisfactorily. I now recommend the manuscript for publication.

·

Basic reporting

No comment.

Experimental design

No comment.

Validity of the findings

No comment.

Comments for the author

Authors addressed all suggestions made by this reviewer. I recommend article should be accepted for publication.

Version 0.1 (original submission)

· · Academic Editor

Minor Revisions

Please pay attention to all the criticisms raised by the referees, especially on how the discussion could be improved if you could comment about some peptides described in high quantities and were not highlighted in the Discussion section so far.

·

Basic reporting

The work was well developed and is clearly written. The authors identified glycoproteins of Haemonchus contortus ligants to galectins of the host and evaluate the putative importance of these proteins to development of vacccines against the parasite.


Line 43: There is not any previous information about "these glyocoproteins".
I think that it is necessary to include information about glyocoproteins before this sentence.

Line 47: please, include references.
Line 48 : add the expression "the expression of" between "upregulate" and "two"

Line 214: add an space in "andtranslation"

Experimental design

Line 168: Was it used a ovine database to identify host protein contamination? I think it is an important control for a better analysis.

Validity of the findings

Line 217: Are actin myosin and collagen from host or from parasite?

·

Basic reporting

Manuscript is generally well written, although there are grammatical corrections needed and English should be reviewed to improve the paper. There are occasional misspellings and grammatical errors throughout the paper. Manuscript does not require expert English editing, but a thorough reading of the final draft is necessary. I have added some comments and suggestions to authors in "General comments for the author".

Experimental design

Experiments were well designed and conducted properly.

Validity of the findings

The study describes L4 and adult H. contortus-derived proteins interacting with host galectins using a pull-down assay followed by LC-MS/MS.

This manuscript contains very useful information for the field and findings described by authors might have implications for the design of new and efficient anti-H. contortus vaccines.

I appreciate authors already had deposited raw LC-MS/MS data on a public repository.

The main weakness is that authors failed to discuss some proteins (such as peptidase S28, alpha beta hydrolize) which were identified in high abundance.

I have added some comments and suggestions to authors in "General comments for the author".

Comments for the author

The manuscript "Proteomic identification of galectin-11 and 14 ligands from Haemonchus contortus” uses pull-down assay followed by spectrometry-based proteomics approach to identify H. contortus proteins that interact with host galectin-11 and galectin-14. The experimental design is well conducted and manuscript is generally well written, although there are grammatical corrections needed and English should be reviewed to improve paper. Some points need to be corrected/clarified before I can recommend it for publication. These are my suggestions:


Major comments:

Material and Methods:
Line 132-133: how much total protein was diluted and applied on column?

Results and Discussion
Composition of LGALS-11 and LGAL-14 ligands:
Line 223-226: in addition to glycan-independent interaction, authors should discuss other types of glycosylation already describe in other organism that are not predicted by tools used in this study.
Please see: Schachter, H. Protein glycosylation lessons from Caenorhabditis elegans. Current Opinion in Structural Biology, 14(5), 607-616, 2004

Protease and phosphatase ligands
Line 256-259: missing discussion about the role(s) of metalloproteases into parasite-host relationship.
Why did not authors discuss in depth the identification and the putative role(s) of “peptidase S28” and “alpha beta hydrolase fold 1”?
According to peptide spectrum matches on Table 1, “peptidase S28” and “alpha beta hydrolase fold 1” appear as the most abundant proteins identified in this study.
Similarly, would suggest to analyze homology/similarity among entries identified in this study and classified as peptidase S28 (W6NE18, W6NFT9, W6NG90).

Tables and Figures
Table 1: I would like to suggest authors to include NSAF values for identified proteins.
I would like to suggest changing “Host-parasite interaction” classification, since all proteins identified could be classified in this class (same for Figure 4).

Table 2 should be removed or presented as Supplemental information, since information contained (presence in adults) is also presented at Table 1 (Groups 2 and 4). Signal peptide and glycosylation predictions should be incorporated at Table 1. I would like to suggest authors to include the number of putative N- and/or O-glycosylation sites for each entry. Description of molecular mass for each protein at Table 1 also would be useful to analyze this information along with electrophoretic profile presented in Figure 2.

Figure 1: Authors did a great job on this picture; however its content (pull down concept) is very basic. I would like to suggest to remove or include as Supplemental information.
Figure 1 legend: “L4 larvae” instead “larval”.
“…Mascot (Perkins et al., 1999) and searched against NCBI protein database”. In Material and Methods section authors described Swiss-Prot database was used.
Figure 2. I would like suggest to authors rephrase title.
Also, I would like to suggest authors to include SDS-PAGE (and results of identified proteins) from control resin (no galectin bound Sepharose). These could be presented as Supplemental information.
Figure 5. Relabel “14_Adult” and “11_Adult”


Minor comments:
Title: “galectin-11” instead “Galectin-11”.

Introduction
Line 46: “mediates”.
Line 47: “self and non-self glycans”.
Line 48: missing a bracket after “LGALS-14”.
Line 49: if possible, please describe species names for Ostertagia and Cooperia.
Line 58-59: missing reference for this sentence.
Line 66: would suggest to replace second “LGALS-11” by a pronoun.

Material and Methods:
Line 84: “normal saline” instead “biological saline” (please review through manuscript).
Line 87: please describe which fungicide was used.
Line 114: I guess “SDS-PAGE” was left by mistake.
Line 119: “Sepharose” (please review through manuscript)
Line 126: Does not blocking buffer contain ethanolamine/glycine or another molecule to block reactive groups on resin?
Line 127: is not clear if both Tris-HCl and sodium acetate buffers contain NaCl.
General for buffer description: please standardize description (put in order molarity, compound, pH, etc...spaces after concentration, etc… try to use IUPAC recommendations).
Line 137: “galactose”
Line 144: “Mass spectrometry”
Line 161: “data-dependent acquisition”.
Line 167-170: Please rephrase this sentence.
Line 172: “were included”.
Line 174: “peptide”
Line 180: please remove “scores” or replace by "values.
Line 182: remove quotation mark after “interactome”.
Line 189: “signal”

Results and Discussion
Line 194: “Identification of proteins of H. contortus that interacting with LGALS-11 and LGALS-14” does not sound good. I would suggest to replace by “Identification of proteins from H. contortus that interact with LGALS-11 and LGALS-14” or “Identification of proteins from H. contortus interacting with LGALS-11 and LGALS-14”.
Line 195 and 196: Please rephrase this sentence.
Line 201: “…were eluted with elution buffer-containing galactose”.
Line 202: “a control”.
Line 214: “and translation”.
Line 243-248: spell out protein names using lowercase letter (please review manuscript).
Did you identified a von Willebrand factor-like protein or a protein containing a von Willebrand factor-like domain?
Line 250: “were identified in adults” instead “in adults were identified”.
Line 251: “omic” (missing a quotation mark).
Line 266: “The von Willebrand”.
Line 279: “identified interacting with host galectins”.
Line 286: reference (Yatsuda et al., 2003) is italicized.

Conclusion
Line 309: “antihelminthic”.
Table 1: “mass spectrometry” instead “mass spectroscopy”.

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