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  6. Journal Club: Structural Basis for Altered Activity of M- and H-Isozyme Forms of Human Lactate Dehydrogenase

20FA_CH428A-Advanced Biochemistry

Journal Club: Structural Basis for Altered Activity of M- and H-Isozyme Forms of Human Lactate Dehydrogenase


Lactate dehydrogenase (LDH) interconverts

pyruvate and lactate with concomitant

interconversion of NADHand NAD1. Although crystal

structures of a variety of LDH have previously

been described, a notable absence has been any of

the three known human forms of this glycolytic

enzyme. We have nowdetermined the crystal structures

of two isoforms of human LDH—the M form,

predominantly found in muscle; and the H form,

found mainly in cardiac muscle. Both structures

have been crystallized as ternary complexes in the

presence of the NADH cofactor and oxamate, a

substrate-like inhibitor. Although each of these isoforms

has different kinetic properties, the domain

structure, subunit association, and active-site regions

are indistinguishable between the two structures.

The pKa that governs the KM for pyruvate for

the two isozymes is found to differ by about 0.94 pH

units, consistent with variation in pKa of the activesite

histidine. The close similarity of these crystal

structures suggests the distinctive activity of these

enzyme isoforms is likely to result directly from

variation of charged surface residues peripheral to

the active site, a hypothesis supported by electrostatic

calculations basedoneachstructure.

Click Read_et_al-2001-Proteins-_Structure,_Function,_and_Bioinformatics.pdf link to view the file.
◄ Structural Determination of Proteins
Tutorial on 2D-NMR ►

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