Ramakrishnan Rajasekhar
Department of Pediatrics, Columbia University College of Physicians and Surgeons, New York NY 10032, USA.
Metabolism. 2007 Nov;56(11):1550-60. doi: 10.1016/j.metabol.2007.06.023.
In a constant infusion study of a mass isotope of leucine, two alternative equations are commonly available to calculate amino acid oxidation rate and, thence, whole-body protein synthesis. One, developed by Matthews et al (Am J Physiol Endocrinol Metab. 1980;238:E473-E479), is shown here to require assuming a tracee steady state (TSS), namely, that tracee (unlabeled) amino acid concentrations and fluxes (rates of oxidation and incorporation into protein) are unaltered compared with the preinfusion state. The other, developed by Garlick and coworkers (Melville et al, Metabolism 1989;38:248-255), stems from a protein steady state (PSS) assumption, namely, that protein synthesis is unaffected by the tracer infusion. We derive here a simple expression for the relative difference in whole-body protein synthesis computed from the two assumptions, and a simple test of the validity of TSS in the form of an equality that must be satisfied by plasma measurements at all times. We also propose two experiments to discriminate between the two assumptions. Theoretical reasons and experimental evidence from the literature are offered to support PSS. The two assumptions result in different expressions for fractional synthetic rates (FSRs) of individual or organ proteins-TSS requires the use of tracer-to-tracee ratios and PSS the use of enrichments. An expression is derived here for the relative difference in FSR with TSS vs PSS. For both whole-body synthesis and for FSR, the TSS assumption consistently results in an underestimate, the relative bias roughly equal to the precursor amino acid enrichment.
在一项亮氨酸大量同位素的持续输注研究中,通常有两个可供选择的方程式来计算氨基酸氧化率,进而计算全身蛋白质合成。其中一个由马修斯等人(《美国生理学杂志:内分泌与代谢》,1980年;238卷:E473 - E479)提出,此处表明该方程式需要假设示踪剂稳态(TSS),即与输注前状态相比,示踪剂(未标记)氨基酸浓度和通量(氧化率和掺入蛋白质的速率)未发生改变。另一个由加利克及其同事(梅尔维尔等人,《新陈代谢》,1989年;38卷:248 - 255)提出,源于蛋白质稳态(PSS)假设,即蛋白质合成不受示踪剂输注的影响。我们在此推导出一个关于根据这两个假设计算出的全身蛋白质合成相对差异的简单表达式,以及一种以等式形式对TSS有效性进行简单检验的方法,该等式必须在所有时间点的血浆测量中都得到满足。我们还提出了两个实验来区分这两个假设。文中提供了理论依据和来自文献的实验证据来支持PSS。这两个假设导致个体或器官蛋白质的分数合成率(FSR)有不同的表达式——TSS需要使用示踪剂与示踪剂前体的比率,而PSS需要使用富集度。在此推导出TSS与PSS相比FSR相对差异的表达式。对于全身合成和FSR而言,TSS假设始终导致低估,相对偏差大致等于前体氨基酸富集度。