Discrimination of cardiac subcellular creatine kinase fluxes by NMR spectroscopy: a new method of analysis.

Abstract:

:A challenge in the understanding of creatine kinase (CK) fluxes reflected by NMR magnetization transfer in the perfused rat heart is the choice of a kinetic model of analysis. The complexity of the energetic pathways, due to the presence of adenosine triphosphate (ATP)-inorganic phosphate (Pi) exchange, of metabolite compartmentation and of subcellular localization of CK isozymes cannot be resolve from the sole information obtained from a single NMR protocol. To analyze multicompartment exchanges, we propose a new strategy based on the simultaneous analysis of four inversion transfer protocols. The time course of ATP and Phosphocreatine (PCr) magnetizations computed from the McConnell equations were adjusted to their experimental value for exchange networks of increasing complexity (up to six metabolite pools). Exchange schemes were selected by the quality of their fit and their consistency with data from other sources: the size of mitochondrial pools and the ATP synthesis flux. The consideration of ATP-Pi exchange and of ATP compartmentation were insufficient to describe the data. The most appropriate exchange scheme in our normoxic heart involved the discrimination of three specific CK activities (cytosolic, mitochondrial, and close to ATPases). At the present level of heart contractility, the energy is transferred from mitochondria to myofibrils mainly by PCr.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Joubert F,Hoerter JA,Mazet JL

doi

10.1016/S0006-3495(01)75940-1

subject

Has Abstract

pub_date

2001-12-01 00:00:00

pages

2995-3004

issue

6

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(01)75940-1

journal_volume

81

pub_type

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