不同水生脊椎动物的血清与黏液蛋白图谱分析

ANALYSIS OF PROTEIN PROFILES IN SERUM AND MUCUS FROM DIFFERENT AQUATIC VERTEBRATES

因脊椎动物的血清与黏液中含有大量蛋白质及其他生物活性物质,而具有多种生理、生化与抗病防御功能。为了解水生脊椎动物血清与黏液中所含主要蛋白成分及其异同,我们分别从中华鲟(Chinese stur-geon Aclpenser sinensis grdy)、鲫鱼(Crucian carp Carassius auratus)、草鱼(Grass carp Ctenopharyngodonidellus)、赤点石斑鱼(Red-spotted grouper Epinephelus akaara)、青石斑鱼(Banded grouper Epinephelusawoara)、狗鱼(Pikes Esox reicherti)、江豚(Finless porpoise Neophocaena phocaenoides)、黄颡鱼(Yellow catfishPelteobagrus fulvidraco)、鳜鱼(Mandarin fish Siniperca chuatsi)这9种水生脊椎动物中,采集血清和皮肤黏液样本。利用考马斯亮蓝G-250法,测定了不同水生脊椎动物的血清和黏液样品中总蛋白的含量;经SDS-PAGE蛋白电泳,分别获得不同水生脊椎动物血清及黏液蛋白图谱;比较和分析了同种动物血清和黏液、不同动物之间血清与血清或黏液与黏液之间蛋白图谱的差异。结果表明:这些动物的血清蛋白组分有显著相似性,在分子量45—120 kD之间,蛋白条带大量集中分布;除青石斑鱼外,其他8种水生脊椎动物都有分子量约为28 kD和14 kD相同或相近的两条蛋白带。草鱼、鲫鱼、鳜鱼、狗鱼、黄颡鱼和江豚的黏液蛋白,除了都含有分子量大小约为45 kD的蛋白条带外,其他无显著相似性。对同种水生脊椎动物的血清与黏液蛋白比较,虽然黄颡鱼的血清与黏液中分子量相近的蛋白带最多,也仅约为38%,可见其蛋白带的种类和含量都存在显著差异。

Aquatic vertebrates live in the water environment with complex and changeable physiological and biochemical reactions.The range of roles which blood plays is very large,such as regulating the physiological and biochemical reactions,maintaining the balance of the organism,ensuring various life activities in an orderly manner.Mucus affords a platform for communication between organism and water environment which is considered as a highly multifunctional material owing to the proposed roles in excretion,respiration,ionic and osmotic regulation,communication,reproduction and protection.The serum and mucus of vertebrates contain a large number of protein components and other biologically active substance as the main bearers of life,which have varieties of function in physiological and biochemical and resistance to pathogenic microorganisms.In order to understand the similarities and differences of the major protein components contained in serum and mucus of aquatic vertebrates,we collected the samples of serum and cutaneous mucus from nine different aquatic animals,such as Chinese sturgeon（Aclpenser sinensis grdy）,crucian carp（Carassius auratus）,grass carp（Ctenopharyngodon idellus）,red-spotted grouper（Epinephelus akaara）,banded grouper（Epinephelus awoara）,pikes（Esox reicherti）,finless porpoise（Neophocaena phocaenoides）,yellow catfish（Pelteobagrus fulvidraco） and mandarin fish（Siniperca chuatsi）.The concentration of serum and mucus protein was determined by coomassie brilliant blue G-250 method and the protein patterns of serum and mucus from the different aquatic vertebrates were observed by SDS-PAGE.The results showed that the range of serum protein concentration was from 17.88 to 31.60 mg/mL and the mucus was from 1.54 to 3.90 mg/mL.The concentration of serum protein from finless porpoise was the highest up to 68.19 mg/mL which was more than twice the red-spotted grouper having the highest serum protein concentration among eight species of fish.On the other hand,the concentration of mucus protein from finless porpoise was the lowest,only 0.45 mg/mL.Therefore,the finless porpoise possessed the largest ratio of protein content in serum and mucus.The protein patterns of serum from the nine aquatic vertebrates might have significant similarities.There were multiple protein bands distribution in the area between molecular weight of 45 kD and 120 kD.Two bands whose molecular weights were approximately 28 kD and 14 kD appeared in the SDS-PAGE protein patterns of serum from the eight aquatic vertebrates,except banded grouper.Other than the band with molecular weight of about 45 kD,no significant similarity was found among the protein patterns of mucus from crucian carp,yellow catfish,mandarin fish,pikes,grass carp and finless porpoise.It showed significant differences in the protein patterns of serum and mucus from same species of aquatic vertebrates.Among five aquatic vertebrates,pikes,mandarin fish,yellow catfish,crucian carp and finless porpoise,the protein patterns of serum and mucus from yellow catfish had the most similarity,and only 38%.The study may provide new material for the further researching on the features and similarities and differences of protein components in serum and mucus from different aquatic vertebrates.