Identification of oxidative post-translational modification of serum albumin in patients with idiopathic pulmonary arterial hypertension and pulmonary hypertension of sickle cell anemia.

Pulmonary hypertension (PH) in sickle cell anemia (SCA) is characterized by decreased nitric oxide bioavailability that might, in part, be related to oxidative stress. Oxidative post-translational modifications of plasma proteins may serve as hallmarks of disease severity and could result in altered protein function and structure. We hypothesized that serum albumin in patients with PH of SCA undergoes oxidative post-translational modification and that this modification may reflect important mediators of disease pathogenesis that are common to both idiopathic pulmonary arterial hypertension (IPAH) and PH of SCA. To explore this hypothesis, we studied albumin purified from the plasma of patients in four subject groups: SCA and PH, SCA steady-state without PH, IPAH, and normal volunteers. Purified albumin was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS). Using MALDI-TOFMS, we identified that an ion corresponding to a malondialdehyde (MDA)-modified albumin peptide was differentially present in patients with IPAH and PH of SCA. These results were confirmed by dot-blotting and Western analysis. We localized the site of MDA modification to albumin residue K159 using LC/MS/MS. Thus, we have identified an MDA modification of serum albumin that appears to be a common link between PH of SCA and IPAH. This finding supports the notion that oxidative stress modulates the pathogenesis of PH of SCA and suggests that this and other post-translational modifications may be important biomarkers of disease.