PXD014575

PXD014575 is an original dataset announced via ProteomeXchange.

Title	Protein-level evidence of novel β-type hemocyanin and heterogeneous subunit usage in the Pacific whiteleg shrimp, Litopenaeus vannamei
Description	The functional diversity of crustacean hemocyanins is broad, encompassing O2 delivery, innate immune response, metabolite storage, and osmolyte balance, all in a heterogeneous protein structure. As such, the sequence diversity of this class of proteins and its subunit composition are the focus of many studies on crustacean adaptation to environmental challenges. Recent transcriptomic and genomic sequencing on the Pacific whiteleg shrimp Litopenaeus vannamei has identified unique isoforms of hemocyanin including an ancestral β-type subunit thought to be lost in penaeid shrimp. However, it is unknown the degree to which these isoforms are translated as proteins, and whether they differ in function. The present study uses proteomic approaches to characterize the protein-level abundance and organization of these hemocyanin isoforms within their native oligomeric structures. Fractions of each hemocyanin oligomeric form were purified by size-exclusion high performance liquid chromatography for identification of subunit isoforms using tandem mass spectrometry at <1% protein false discovery rate. Relative abundances of hemocyanin oligomers and monomeric subunits from hemolymph and fractions were also quantified by polyacrylamide gel electrophoresis with and without denaturation for comparison of relative abundance. Ten hemocyanin isoforms were identified by tandem mass spectrometry in both hemocyanin oligomer fractions including a single small subunit, eight large subunits, and the first protein-level evidence of a β-type subunit in penaeid shrimp. Hemocyanin subunits were organized primarily as hexamers (95-99% relative abundance) as opposed to dodecamers. Hexamers utilized a significantly higher ratio (2.05:1) of small subunit to large subunit compared to dodecamers (1.04:1), and the relative abundances of the large subunit isoforms was dominated by HcL1 in both fractions. The ability to distinguish and quantify hemocyanin isoforms within oligomeric structures will aid future studies linking hemocyanin genes to function and physiology as well as offer insight into the evolutionary history of crustaceans.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_06:59:26.357.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jason Wang
SpeciesList	scientific name: Penaeus vannamei; NCBI TaxID: 6689;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue; deamidated residue
Instrument	TripleTOF 5600

Revision	Datetime	Status	ChangeLog Entry
0	2019-07-11 07:59:11	ID requested
1	2020-02-07 03:23:02	announced
2	2020-02-07 04:24:28	announced	2020-02-07: Updated publication reference for DOI(s): 10.3389/fmars.2019.00687.
⏵ 3	2023-11-14 06:59:26	announced	2023-11-14: Updated project metadata.

Dataset with its publication pending

submitter keyword: proteomics, subunit heterogeneity, mass spectrometry,hemocyanin, crustacean

Karen Gray Burnett
contact affiliation	Biology Department, College of Charleston, SC, USA
contact email	burnettk@cofc.edu
lab head
Jason Wang
contact affiliation	University of Southern California
contact email	jasonwang103@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD014575
     1. Label: PRIDE project
     2. Name: Protein-level evidence of novel β-type hemocyanin and heterogeneous subunit usage in the Pacific whiteleg shrimp, Litopenaeus vannamei