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Conformational stability and in vitro bioactivity of porcine luteinizing hormone
Journal article   Peer reviewed

Conformational stability and in vitro bioactivity of porcine luteinizing hormone

Tanya Burova, François Lecompte, Colette Galet, Françoise Monsallier, Suzanne Delpech, Thomas Haertlé and Yves Combarnous
Molecular and cellular endocrinology, Vol.176(1), pp.129-134
05/15/2001
DOI: 10.1016/S0303-7207(01)00447-6
PMID: 11369452

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Abstract

Temperature-dependent dissociation of porcine luteinizing hormone (pLH) and of two of its glycoforms was studied by a combination of SDS-PAGE and micro-scale size-exclusion HPLC in parallel with the study of co-operative folding by high-sensitivity differential scanning calorimetry (HS-DSC). The transition temperature of dissociation of pLH at pH 7.0 as quantified by SDS-PAGE, HPLC and residual activity in radioreceptor assay was found to match exactly the transition temperature of its unfolding as measured by HS-DSC. Free α- and β-subunits did not exhibit any unfolding transition in the same conditions. The microcalorimetric data for two pLH isoforms exhibiting different glycosylations were identical to those of a preparation of non-separated isoforms. It is concluded that: (a) free subunits exhibit no co-operative folding (i.e. no stable three-dimensional structure) and co-operative folding occurs only in αβ heterodimers; (b) the co-operative folding is responsible for the stability of the association of subunits; and (c) the heterogeneity of carbohydrate chains does not affect the stability of folding and association of subunits. The fastening of the ‘seat-belt’ of the β-subunit embracing the α-subunit by the Cysβ26-β110 disulfide bridge had been postulated to play a role in the preservation of the dimeric structure of gonadotropins. The present work shows that dissociation of subunits is directly related to their loss of common co-operative folding.
Differential scanning calorimetry Porcine luteinizing hormone Quaternary structure stability Receptor binding

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