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Adiponectin secretion from cardiomyocytes produces canonical multimers and partial co-localization with calsequestrin in junctional SR

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Abstract

Adiponectin (ADN) is an abundant protein in serum, secreted by adipocytes, that acts as a signal for fat metabolism. It is marked by a complex molecular structure that results from processes within the secretory pathway, producing a canonical set of multimers. ADN may also be secreted from cardiomyocytes, where a unique sarcomeric endoplasmic/sarcoplasmic reticulum (ER/SR) substructure has been characterized primarily for its Ca handling. We expressed ADN in cultured primary adult cardiomyocytes and nonmuscle (COS) cells. After 48 h of ADN expression by adenovirus treatment, roughly half of synthesized ADN was secreted from cardiomyocytes, and half was still in-transit within inner membrane compartments, similar to COS cells. Cardiomyocytes and COS cells both produced ADN in the three canonical forms: trimers, hexamers, and 18-mers. Higher rates of secretion occurred for higher-molecular weight multimers, especially 18-mers. The highest levels of ADN protein, whether in transit or secreted, were present as trimers and hexamers. In nonmuscle cell lines, ADN trafficked through ER and Golgi compartments as expected. In contrast, ADN in primary adult cardiomyocytes populated ER/SR tubules along the edges of sarcomeres that emanated from nuclear surfaces. Prominent co-localization of ADN occurred with calsequestrin, a marker of junctional SR, the Ca2+-release compartment of the cell. The early steps in ADN trafficking re-trace those recently described for newly made junctional SR proteins, involving a nuclear envelope (NE) translocation into SR tubules that are oriented along sarcolemmal transverse (T)-tubules (NEST pathway).

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Abbreviations

ER:

Endoplasmic reticulum

jSR:

Junctional sarcoplasmic reticulum

ADN:

Adiponectin

MOI:

Multiplicity of infection

Pfu:

Plaque-forming units

HMW:

High molecular weight

MMW:

Medium molecular weight

LMW:

Low molecular weight

ADN-WT:

Wild-type (human) adiponectin

ADN-Flag:

Flag-tagged adiponectin

native-PAGE:

Native polyacrylamide gel electrophoresis

BFA:

Brefeldin A

NEST pathway:

Nuclear envelope to SR along T-tubules pathway

z-tubules:

SR tubules that traffic newly made proteins roughly aligned with the sarcomeric z-line and T-tubules

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Acknowledgements

This work was supported by an Incubator Grant 1-77311 from the Office of Vice President for Research, Wayne State University.

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Authors and Affiliations

  1. Department of Physiology, Wayne State University, Detroit, MI, 48201, USA

    Joanna Solarewicz, Amanda Manly, Stephanie Kokoszka, Naama Sleiman & Steven Cala

  2. Department of Pathology, Wayne State University, Detroit, MI, 48201, USA

    Todd Leff

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  1. Joanna Solarewicz
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Correspondence to Steven Cala.

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Solarewicz, J., Manly, A., Kokoszka, S. et al. Adiponectin secretion from cardiomyocytes produces canonical multimers and partial co-localization with calsequestrin in junctional SR. Mol Cell Biochem 457, 201–214 (2019). https://doi.org/10.1007/s11010-019-03524-9

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