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首页> 美国卫生研究院文献>Journal of Animal Science >PSI-28 Genomic determinants of alkaline phosphatase catalytic affinity along the intestinal longitudinal axis of weanling pigs.
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PSI-28 Genomic determinants of alkaline phosphatase catalytic affinity along the intestinal longitudinal axis of weanling pigs.

机译：PSI-28断奶仔猪肠道纵轴上碱性磷酸酶催化亲和力的基因组决定因素。

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摘要

Alkaline phosphatases (AP) play a pivotal role in protecting intestinal health against pathogenic bacterial toxins through dephosphorylation of endotoxin LPS lipid moiety and other emblematic members of pathogen-associated molecular patterns such as ATP. This study was conducted to elucidate the genomic determinants of alkaline phosphatase catalytic affinity along the intestinal longitudinal axis of weanling pigs. Herein, jejunal, ileal, cecal and colonic segmental samples were collected from 8 weanling barrows fed a corn and SBM-based diet, with an average final BW of 10.95 (SE=0.68) kg. These gut tissues were homogenized for AP kinetics characterization using p-nitrophenyl phosphate as a substrate at pH 7.4 and 37 ºC for 30 min (parameter estimates±SE, P2=0.33 – 0.69, n=40). The AP affinity (Km, µM) in the jejunum (27.40 ± 9.66) and the colon (30.01 ± 12.42) was lower (PVmax, nmol·mg proteinPSus scrofa Ver. 11.1) further identified 5 AP genes, including 3 intestinal-like AP isomer genes located in the chromosome 15, and 2 tissue-non-specific AP isomer genes in the chromosome 6, resulting in 5 AP isozymes that are all likely expressed in the intestine. The predicted 3-D protein structure models for the 5 AP isomers have revealed that although the catalytic site architectures, including the heavy metal-binding sites, are highly conserved, the mapped N-glycosylation sites are very variable among the 5 AP isomers. Thus, differences in these AP isomer distribution and the different N-glycosylation sites are likely the major genomic determinants affecting the alkaline phosphatase catalytic affinity along the intestinal longitudinal axis and gut health status of weanling pigs.
机译：碱性磷酸酶（AP）通过内毒素LPS脂质部分和病原体相关分子模式的其他标志性成员（例如ATP）的去磷酸化，在保护肠道健康免受病原性细菌毒素的侵害中发挥关键作用。进行该研究以阐明沿断奶猪肠纵轴的碱性磷酸酶催化亲和力的基因组决定因素。在这里，从饲喂玉米和基于SBM的日粮的8个断奶公猪收集空肠，回肠，盲肠和结肠的节段样本，平均最终体重为10.95（SE = 0.68）kg。使用磷酸对硝基苯酯作为底物，在pH 7.4和37ºC下30分钟将这些肠组织匀浆以进行AP动力学表征（参数估计值±SE，P2 = 0.33 – 0.69，n = 40）。空肠（27.40±9.66）和结肠（30.01±12.42）中的AP亲和力（Km，µM）较低（PVmax，nmol·mg蛋白PSus scrofa Ver.11.1）进一步鉴定了5个AP基因，包括3个肠样AP位于15号染色体上的异构体基因和6号染色体上的2个组织非特异性AP异构体基因，导致5种AP同工酶都可能在肠中表达。预测的5个AP异构体的3-D蛋白质结构模型表明，尽管催化位点结构（包括重金属结合位点）是高度保守的，但映射的N-糖基化位点在5个AP异构体之间变化很大。因此，这些AP异构体分布的差异和不同的N-糖基化位点可能是影响沿断奶仔猪肠道纵轴和肠道健康状况的碱性磷酸酶催化亲和力的主要基因组决定因素。

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