A 35-kDa polypeptide of the crystalline lens in the common frog: its biochemical properties, tissue specificity and appearance in the developmental process

• Published in: Ontogenez Vol. 21; no. 5; p. 487
• Main Authors: Simirskiĭ, V N, Aleĭnikova, K S, Mikhaĭlov, A T
• Format: Journal Article
• Language: Russian
• Published: Russia (Federation) 01.09.1990
• Subjects: Animals; Crystallins - metabolism; Immunochemistry; Larva - growth & development; Lens, Crystalline - metabolism; Molecular Weight; Morphogenesis - physiology; Organ Specificity - physiology; Peptides - analysis; Peptides - isolation & purification; Peptides - metabolism; Rana temporaria - growth & development
• ISSN: 0475-1450

The vertebrate lens contains so-called taxon-specific water-soluble proteins. One of them is p-crystallin with a molecular weight of 35 kDa characteristic of Ranidae family. We have identified a polypeptide with a molecular weight of 35 kDa in the eye lens of Rana temporaria which: (1) can be extracted from the lens by aqueous salt solutions, (2) has a molecular mass of 36.1 +/- 0.4 kDa (by SDS-electrophoresis) and 37 kDa (by gel filtration), (3) is heterogeneous in terms of isoelectric point (pI 6.5-8.0), (4) binds to heparin-agarose, (5) denatures in response to freezing-thawing, lyophilization and in solutions with low ionic strength. Thus, major biochemical parameters of this polypeptide differ from that of amphibian alpha, beta- and gamma-crystallins. In addition to lens, 35 kDa polypeptide was detected by immunoelectroblotting in retina, testes, liver, kidney, spleen, stomach, intestine and lungs. Its level (as percentage of water-soluble protein) is 1.1 +/- 1.4% in the lens, 1.6 +/- 0.7% in retina. 0.05% in testes and liver and 0.01% or less in other organs. Thus, despite its wide tissue distribution, 53 kDa polypeptide is expressed predominantly in lens and retina. We studied the time-course of appearance and accumulation of this polypeptide in tissues where it is expressed at high or low levels. 35 kDa polypeptide was detected for the first time during larval development: (1) in the lens (some time after the mouth opening; stages 33-34 according to Dabagian and Sleptsova, 1975), (2) in the retina (by the time of anus opening; stages 36-37), (3) in the liver (at the stage of elongated hind limb bud; stages 40-41). Definitive expression level of this protein was achieved in the lens by the beginning of metamorphosis and in the retina and liver during first months of development. Hence, during the whole period of larval development 35 kDa polypeptide content of the lens exceeds that of retina or liver. A more substantial evidence is required to confirm the identity of studied polypeptide with rho-crystallin.