Affimers targeting proteins in the cardiomyocyte Z-disc: Novel tools that improve imaging of heart tissue: Frontiers in Cardiovascular Medicine

F. Parker, A.A.S. Tang, B. Rogers, G. Carrington, C. dos Remedios, A. Li, D. Tomlinson, M. Peckham

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Dilated Cardiomyopathy is a common form of heart failure. Determining how this disease affects the structure and organization of cardiomyocytes in the human heart is important in understanding how the heart becomes less effective at contraction. Here we isolated and characterised Affimers (small non-antibody binding proteins) to Z-disc proteins ACTN2 (α-actinin-2), ZASP (also known as LIM domain binding protein 3 or LDB3) and the N-terminal region of the giant protein titin (TTN Z1-Z2). These proteins are known to localise in both the sarcomere Z-discs and the transitional junctions, found close to the intercalated discs that connect adjacent cardiomyocytes. We use cryosections of left ventricles from two patients diagnosed with end-stage Dilated Cardiomyopathy who underwent Orthotopic Heart Transplantation and were whole genome sequenced. We describe how Affimers substantially improve the resolution achieved by confocal and STED microscopy compared to conventional antibodies. We quantified the expression of ACTN2, ZASP and TTN proteins in two patients with dilated cardiomyopathy and compared them with a sex- and age-matched healthy donor. The small size of the Affimer reagents, combined with a small linkage error (the distance from the epitope to the dye label covalently bound to the Affimer) revealed new structural details in Z-discs and intercalated discs in the failing samples. Affimers are thus useful for analysis of changes to cardiomyocyte structure and organisation in diseased hearts. Copyright © 2023 Parker, Tang, Rogers, Carrington, dos Remedios, Li, Tomlinson and Peckham.
Original languageEnglish
JournalFrontiers in Cardiovascular Medicine
Volume10
DOIs
Publication statusPublished - 2023

Keywords

  • affimer targeting protein
  • alpha actinin 2
  • ccstem28
  • complementary DNA
  • cysteine
  • desmoglein 2
  • ea 53
  • epitope
  • immunoglobulin G
  • LIM domain binding protein 3
  • monoclonal antibody
  • monoclonal antibody CCSTEM28
  • monoclonal antibody EA 53
  • monoclonal antibody Z1Z2
  • myosin binding protein C
  • n terminal region of the giant protein titin
  • protein
  • unclassified drug
  • Z disc protein
  • z1z2
  • ZASP protein
  • adult
  • alphafold modelling
  • antibody labelling
  • Article
  • case report
  • centrifugation
  • clinical article
  • confocal microscopy
  • congestive cardiomyopathy
  • controlled study
  • coronary artery blood flow
  • cryosections
  • crystal structure
  • downstream processing
  • dye label covalently bound
  • enzyme linked immunosorbent assay
  • epitope mapping
  • expression vector
  • female
  • frameshift mutation
  • heart muscle contractility
  • heart tissue
  • human
  • human cell
  • human tissue
  • imaging
  • intercalated disc
  • linkage analysis
  • male
  • nonsense mutation
  • orthotopic heart transplantation
  • phage display
  • protein expression
  • protein structure
  • protein targeting
  • sarcomere
  • sarcomere length
  • sequence analysis
  • size exclusion chromatography
  • stimulated emission depletion microscopy
  • stop codon
  • super resolution microscopy
  • tissue penetration
  • transitional junction
  • whole genome sequencing
  • z disc width
  • Affimer
  • cardiac actinin
  • Dilated Cardiomyopathy
  • fluorescence microscopy
  • Z-disc
  • ZASP and the N-terminal region of titin (TTN)

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