Cross-species mechanical fingerprinting of cardiac myosin binding protein-C

Árpád Karsai, Miklós S Z Kellermayer, Samantha - Harris

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Cardiac myosin binding protein-C (cMyBP-C) is a member of the immunoglobulin (Ig) superfamily of proteins and consists of 8 Ig- and 3 fibronectin III (FNIII)-like domains along with a unique regulatory sequence referred to as the MyBP-C motif or M-domain. We previously used atomic force microscopy to investigate the mechanical properties of murine cMyBP-C expressed using a baculovirus/insect cell expression system. Here, we investigate whether the mechanical properties of cMyBP-C are conserved across species by using atomic force microscopy to manipulate recombinant human cMyBP-C and native cMyBP-C purified from bovine heart. Force versus extension data obtained in velocity-clamp experiments showed that the mechanical response of the human recombinant protein was remarkably similar to that of the bovine native cMyBP-C. Ig/Fn-like domain unfolding events occurred in a hierarchical fashion across a threefold range of forces starting at relatively low forces of ∼50 pN and ending with the unfolding of the highest stability domains at ∼180 pN. Force-extension traces were also frequently marked by the appearance of anomalous force drops suggestive of additional mechanical complexity such as structural coupling among domains. Both recombinant and native cMyBP-C exhibited a prominent segment ∼100 nm-long that could be stretched by forces <50 pN before the unfolding of Ig- and FN-like domains. Combined with our previous observations of mouse cMyBP-C, these results establish that although the response of cMyBP-C to mechanical load displays a complex pattern, it is highly conserved across species.

Original languageEnglish (US)
Pages (from-to)2465-2475
Number of pages11
JournalBiophysical Journal
Volume104
Issue number11
DOIs
StatePublished - Jun 4 2013
Externally publishedYes

Fingerprint

Cardiac Myosins
Atomic Force Microscopy
Immunoglobulins
myosin-binding protein C
Baculoviridae
Fibronectins
Recombinant Proteins
Insects

ASJC Scopus subject areas

  • Biophysics

Cite this

Cross-species mechanical fingerprinting of cardiac myosin binding protein-C. / Karsai, Árpád; Kellermayer, Miklós S Z; Harris, Samantha -.

In: Biophysical Journal, Vol. 104, No. 11, 04.06.2013, p. 2465-2475.

Research output: Contribution to journalArticle

Karsai, Árpád ; Kellermayer, Miklós S Z ; Harris, Samantha -. / Cross-species mechanical fingerprinting of cardiac myosin binding protein-C. In: Biophysical Journal. 2013 ; Vol. 104, No. 11. pp. 2465-2475.
@article{00d7592860b64abcb81974bd5faf8978,
title = "Cross-species mechanical fingerprinting of cardiac myosin binding protein-C",
abstract = "Cardiac myosin binding protein-C (cMyBP-C) is a member of the immunoglobulin (Ig) superfamily of proteins and consists of 8 Ig- and 3 fibronectin III (FNIII)-like domains along with a unique regulatory sequence referred to as the MyBP-C motif or M-domain. We previously used atomic force microscopy to investigate the mechanical properties of murine cMyBP-C expressed using a baculovirus/insect cell expression system. Here, we investigate whether the mechanical properties of cMyBP-C are conserved across species by using atomic force microscopy to manipulate recombinant human cMyBP-C and native cMyBP-C purified from bovine heart. Force versus extension data obtained in velocity-clamp experiments showed that the mechanical response of the human recombinant protein was remarkably similar to that of the bovine native cMyBP-C. Ig/Fn-like domain unfolding events occurred in a hierarchical fashion across a threefold range of forces starting at relatively low forces of ∼50 pN and ending with the unfolding of the highest stability domains at ∼180 pN. Force-extension traces were also frequently marked by the appearance of anomalous force drops suggestive of additional mechanical complexity such as structural coupling among domains. Both recombinant and native cMyBP-C exhibited a prominent segment ∼100 nm-long that could be stretched by forces <50 pN before the unfolding of Ig- and FN-like domains. Combined with our previous observations of mouse cMyBP-C, these results establish that although the response of cMyBP-C to mechanical load displays a complex pattern, it is highly conserved across species.",
author = "{\'A}rp{\'a}d Karsai and Kellermayer, {Mikl{\'o}s S Z} and Harris, {Samantha -}",
year = "2013",
month = "6",
day = "4",
doi = "10.1016/j.bpj.2013.04.027",
language = "English (US)",
volume = "104",
pages = "2465--2475",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Biophysical Society",
number = "11",

}

TY - JOUR

T1 - Cross-species mechanical fingerprinting of cardiac myosin binding protein-C

AU - Karsai, Árpád

AU - Kellermayer, Miklós S Z

AU - Harris, Samantha -

PY - 2013/6/4

Y1 - 2013/6/4

N2 - Cardiac myosin binding protein-C (cMyBP-C) is a member of the immunoglobulin (Ig) superfamily of proteins and consists of 8 Ig- and 3 fibronectin III (FNIII)-like domains along with a unique regulatory sequence referred to as the MyBP-C motif or M-domain. We previously used atomic force microscopy to investigate the mechanical properties of murine cMyBP-C expressed using a baculovirus/insect cell expression system. Here, we investigate whether the mechanical properties of cMyBP-C are conserved across species by using atomic force microscopy to manipulate recombinant human cMyBP-C and native cMyBP-C purified from bovine heart. Force versus extension data obtained in velocity-clamp experiments showed that the mechanical response of the human recombinant protein was remarkably similar to that of the bovine native cMyBP-C. Ig/Fn-like domain unfolding events occurred in a hierarchical fashion across a threefold range of forces starting at relatively low forces of ∼50 pN and ending with the unfolding of the highest stability domains at ∼180 pN. Force-extension traces were also frequently marked by the appearance of anomalous force drops suggestive of additional mechanical complexity such as structural coupling among domains. Both recombinant and native cMyBP-C exhibited a prominent segment ∼100 nm-long that could be stretched by forces <50 pN before the unfolding of Ig- and FN-like domains. Combined with our previous observations of mouse cMyBP-C, these results establish that although the response of cMyBP-C to mechanical load displays a complex pattern, it is highly conserved across species.

AB - Cardiac myosin binding protein-C (cMyBP-C) is a member of the immunoglobulin (Ig) superfamily of proteins and consists of 8 Ig- and 3 fibronectin III (FNIII)-like domains along with a unique regulatory sequence referred to as the MyBP-C motif or M-domain. We previously used atomic force microscopy to investigate the mechanical properties of murine cMyBP-C expressed using a baculovirus/insect cell expression system. Here, we investigate whether the mechanical properties of cMyBP-C are conserved across species by using atomic force microscopy to manipulate recombinant human cMyBP-C and native cMyBP-C purified from bovine heart. Force versus extension data obtained in velocity-clamp experiments showed that the mechanical response of the human recombinant protein was remarkably similar to that of the bovine native cMyBP-C. Ig/Fn-like domain unfolding events occurred in a hierarchical fashion across a threefold range of forces starting at relatively low forces of ∼50 pN and ending with the unfolding of the highest stability domains at ∼180 pN. Force-extension traces were also frequently marked by the appearance of anomalous force drops suggestive of additional mechanical complexity such as structural coupling among domains. Both recombinant and native cMyBP-C exhibited a prominent segment ∼100 nm-long that could be stretched by forces <50 pN before the unfolding of Ig- and FN-like domains. Combined with our previous observations of mouse cMyBP-C, these results establish that although the response of cMyBP-C to mechanical load displays a complex pattern, it is highly conserved across species.

UR - http://www.scopus.com/inward/record.url?scp=84878838067&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84878838067&partnerID=8YFLogxK

U2 - 10.1016/j.bpj.2013.04.027

DO - 10.1016/j.bpj.2013.04.027

M3 - Article

C2 - 23746519

AN - SCOPUS:84878838067

VL - 104

SP - 2465

EP - 2475

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 11

ER -