Lysine modification of human serum albumin and its effect on protein conformation and nalidixic acid binding

ABSTRACT The control of protein conformation during translocation through the endoplasmic reticulum is often a bottleneck for heterologous protein production. The core pathway of the oxidative folding machinery includes two conserved proteins: Pdi1p and Ero1p. We increased the dosage of the genes encoding these proteins in the yeast Kluyveromyces lactis and evaluated the secretion of heterologous proteins. KlERO1, an orthologue of Saccharomyces cerevisiae ERO1, was cloned by functional complementation of the ts phenotype of an Scero1 mutant. The expression of KlERO1 was induced by treatment of the cells with dithiothreitol and by overexpression of human serum albumin (HSA), a disulfide bond-rich protein. Duplication of either PDI1 or ERO1 led to a similar increase in HSA yield. Duplication of both genes accelerated the secretion of HSA and improved cell growth rate and yield. Increasing the dosage of KlERO1 did not affect the production of human interleukin 1β, a protein that has no disulfide bridges. The results confirm that the ERO1 genes of S. cerevisiae and K. lactis are functionally similar even though portions of their coding sequence are quite different and the phenotypes of mutants overexpressing the genes differ. The marked effects of KlERO1 copy number on the expression of heterologous proteins with a high number of disulfide bridges suggests that control of KlERO1 and KlPDI1 is important for the production of high levels of heterologous proteins of this type.

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Affinity of rosmarinic acid to human serum albumin and its effect on protein conformation stability

Food Chemistry ◽

10.1016/j.foodchem.2015.06.109 ◽

2016 ◽

Vol 192 ◽

pp. 178-187 ◽

Cited By ~ 69

Author(s):

Xin Peng ◽

Xiangchao Wang ◽

Wei Qi ◽

Rongxin Su ◽

Zhimin He

Keyword(s):

Human Serum Albumin ◽

Serum Albumin ◽

Human Serum ◽

Protein Conformation ◽

Rosmarinic Acid ◽

Conformation Stability

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Betulinic acid binding to human serum albumin: A study of protein conformation and binding affinity

Journal of Photochemistry and Photobiology B Biology ◽

10.1016/j.jphotobiol.2008.09.002 ◽

2009 ◽

Vol 94 (1) ◽

pp. 8-12 ◽

Cited By ~ 88

Author(s):

Rajagopal Subramanyam ◽

Anilkishor Gollapudi ◽

Persis Bonigala ◽

Madhurarekha Chinnaboina ◽

Damu G. Amooru

Keyword(s):

Human Serum Albumin ◽

Serum Albumin ◽

Human Serum ◽

Binding Affinity ◽

Betulinic Acid ◽

Protein Conformation

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Ligand-binding properties of proalbumin Christchurch

Biochemical Journal ◽

10.1042/bj1910281 ◽

1980 ◽

Vol 191 (1) ◽

pp. 281-283 ◽

Cited By ~ 3

Author(s):

R G Reed ◽

T Peters ◽

S O Brennan ◽

R W Carrell

Keyword(s):

Fatty Acids ◽

Human Serum Albumin ◽

Ligand Binding ◽

Serum Albumin ◽

Human Serum ◽

Protein Conformation ◽

Binding Sites ◽

Bromocresol Green ◽

Binding Properties ◽

Normal Albumin

Proalbumin Christchurch, a circulating variant of human serum albumin, is secreted from the liver without cleavage of the hexapeptide situated at the N-terminal end of the peptide chain of proalbumin. We compared ligand-binding properties of proalbumin Christchurch and of normal albumin A from the same individual in order to test the effect of the presence of the hexapeptide. The two albumin forms exhibited similar affinities for palmitate, bilirubin, 8-anilinonaphthalene-1-sulphonate and Bromocresol Green. The patterns of endogenous fatty acids bound to the two forms of albumin were slightly different, although the differences were probably not of physiological significance. From these studies it would appear that the propeptide of proalbumin does not alter the protein conformation in such a way as to alter binding sites for organic anions.

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Spectroscopic investigations on the binding of an iodinated chlorin p6-copper complex to human serum albumin

Photochemical & Photobiological Sciences ◽

10.1039/c7pp00197e ◽

2017 ◽

Vol 16 (12) ◽

pp. 1762-1770 ◽

Cited By ~ 4

Author(s):

P. Sarbadhikary ◽

A. Dube

Keyword(s):

Human Serum Albumin ◽

Serum Albumin ◽

Human Serum ◽

Binding Site ◽

Copper Complex ◽

Protein Conformation ◽

High Affinity ◽

Spectroscopic Investigations ◽

Chlorin P6

An iodinated chlorin p6 copper complex showed high affinity to bind human serum albumin, the binding site was predicted and it was demonstrated that binding did not affect protein conformation.

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Effect of lysine modification on the conformation and indomethacin binding properties of human serum albumin

International Journal of Biological Macromolecules ◽

10.1016/s0141-8130(99)00074-4 ◽

1999 ◽

Vol 26 (2-3) ◽

pp. 173-180 ◽

Cited By ~ 16

Author(s):

Saad Tayyab ◽

Soghra K. Haq ◽

Sabeeha ◽

Mohammad A. Aziz ◽

Mohammad M. Khan ◽

...

Keyword(s):

Human Serum Albumin ◽

Serum Albumin ◽

Human Serum ◽

Binding Properties ◽

Lysine Modification

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Targeting the Nalidixic Acid Binding Site on Human Serum Albumin Through Computational Approach: A Re-Investigation

Biointerface Research in Applied Chemistry ◽

10.33263/briac122.15201525 ◽

2021 ◽

Vol 12 (2) ◽

pp. 1520-1525

Keyword(s):

Molecular Docking ◽

Human Serum Albumin ◽

Serum Albumin ◽

Human Serum ◽

Binding Site ◽

Nalidixic Acid ◽

Dna Supercoiling ◽

Serum Albumins ◽

Binding Preference ◽

Tract Infections

Nalidixic acid (NA) is a quinolone drug used to treat urinary tract infections. It inhibits bacterial gyrase-DNA complex formation, an essential step for DNA supercoiling during bacterial replication. Due to the medical application of this drug, it would be interesting to get insight into its binding mechanism with human serum albumin (HSA), the primary carrier protein in blood circulation. Two reports of NA binding to HSA were published using molecular docking approaches. The first report revealed that the preferred binding site of NA was Site II of serum albumins, while the recent finding predicted Site I of bovine serum albumin (BSA) as the preferred site. Given the high sequence homology between these albumins, it is presumed that the binding preference of this drug should be the same in these proteins. To re-investigate this phenomenon, the interaction of NA with HSA was conducted using AutoDockTool 4.0. The molecular docking results revealed that NA binding preference was at Site I, involving Lys 199 in HSA, due to the formation of more significant contacts. Hence, it is concluded that any variable or parameters in the software should be wisely standardized to minimize the controversial results using different programs.

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