PXD022455

PXD022455 is an original dataset announced via ProteomeXchange.

Title	A function-based determination of the cardiac endoplasmic and sarcoplasmic reticulum proteome
Description	Cardiac sarcoplasmic reticulum (SR) plays a central role in cellular Ca homeostasis, as does endoplasmic reticulum (ER) in the nonmuscle cell. The importance of SR Ca-handling function has led to detailed understanding of Ca-release and re-uptake protein complexes, but relatively little information regarding other known ER functions. We isolated cardiac membranes based upon their ability to efficiently accumulate Ca by the SR,ER-ATPase (SERCA-positive membranes), a process known to produce two characteristic SR membrane fractions: those enriched in known junctional SR markers, and those enriched in proteins originating from classic ER subdomains. Using standard proteomic techniques, we determined the SR proteins in three membrane preparations: 1) the highest-density SERCA-positive microsomes (HighSR); 2) the slightly less dense SERCA-positive microsomes that are constrained by a ryanodine-dependent leak (MedSR); and the original crude cardiac microsomes. Only a third of all crude microsomal proteins in heart were enriched in SERCA-positive membranes at least two fold. This protocol completely excluded known contractile, mitochondrial, sarcolemmal, and other major microsomal proteins, producing a far cleaner preparation of SR. Each SERCA-positive protein was distributed between HighSR and MedSR membrane vesicles in accordance with relative densities of SERCA2a versus ryanodine receptor. The distributions reinforced past findings on the distributions for several of the major known proteins. When combined with values for relative abundance, and enrichment over crude membranes, our protocol conveniently and effectively exposed multiple domains of the cardiac ER/SR. Most of the highly enriched and abundant cardiac SR/ER proteins represent proteins that have received little research attention. The data generate a useful quantitative analysis of intracellular membrane structure and function within cardiomyocytes, and may provide a reliable way of gauging broader changes in cardiac SR.
HostingRepository	PRIDE
AnnounceDate	2024-01-26
AnnouncementXML	Submission_2024-01-26_05:56:17.505.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD022455
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Nicholas Carruthers
SpeciesList	scientific name: Canis familiaris (Dog) (Canis lupus familiaris); NCBI TaxID: 9615;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	LTQ

Revision	Datetime	Status	ChangeLog Entry
0	2020-11-10 07:56:22	ID requested
⏵ 1	2024-01-26 05:56:18	announced

10.6019/PXD022455;

Cala SE, Carruthers NJ, Stemmer PM, Chen Z, Chen X, transport in cardiac microsomes enriches functional sets of ER and SR proteins. Mol Cell Biochem, 479(1):85-98(2024) [pubmed]

10.1007/s11010-023-04708-0;

submitter keyword: proteomics, SERCA, cardiac sarcoplasmic reticulum,cardiac muscle, ryanodine receptor

Steven Cala
contact affiliation	Department of Physiology, Wayne State University
contact email	s.cala@edu.edu
lab head
Nicholas Carruthers
contact affiliation	IEHS
contact email	aj7682@wayne.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD022455
     1. Label: PRIDE project
     2. Name: A function-based determination of the cardiac endoplasmic and sarcoplasmic reticulum proteome