Effect of lysine residue modification of ovine luteinizing hormone by heterobifunctional crosslinking reagent SPDP on subunit-subunit association, receptor binding and biological activity.
The increasing use of heterobifunctional crosslinking agent in the design of hormone-carrier conjugates for selective targeting or inducing immune response against the hormone has prompted us to study the effect of epsilon-NH2 group modification of oLH-subunit, their recombination, immunoreactivity, receptor binding and biological activity. The epsilon-NH2 groups of oLH alpha and oLH beta subunits were modified by using SPDP. The SPDP modified oLH alpha derivatives hybridize to native OLH beta as judged by RP-HPLC analysis. The sequential modification of alpha and beta subunits led to progressive reduction in immunoreactivity and receptor binding activities. The steroidogenic potential of oLH beta.SPDP.alpha oLH recombinant was relatively comparable. The modification of six or more epsilon-NH2 groups in oLH alpha although recombine fully with native oLH beta but failed to react to anti-oLH antibody. Moreover, steroidogenic activity was also abolished. Introduction up to four SPDP groups in oLH alpha compromised immunological and biological activities but further addition of two more SPDP groups completely abolished antibody reactivity, receptor binding and steroidogenic activity indicating the importance of later two -NH2 groups in the receptor recognition and steroidogenic potential.
Singh
V
V
Institute of Self Organising Systems and Biophysics, North-Eastern Hill University, Shillong, India.
Mavila
A K
AK
Alam
A
A
D000818:Animals; D003432:Cross-Linking Reagents; D015292:Glycoprotein Hormones, alpha Subunit / Q000737:chemistry / Q000378:metabolism; D007986:Luteinizing Hormone / Q000737:chemistry* / Q000378:metabolism; D008297:Male; D010902:Pituitary Gland / Q000737:chemistry; D011869:Radioligand Assay; D051381:Rats; D011974:Receptors, LH / Q000378:metabolism; D012756:Sheep; D013053:Spectrophotometry; D013388:Succinimides; D013737:Testis / Q000378:metabolism
D003432:Cross-Linking Reagents; D015292:Glycoprotein Hormones, alpha Subunit; D011974:Receptors, LH; D013388:Succinimides; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate; D007986:Luteinizing Hormone