Refined structure of rat Clara cell 17 kDa protein at 3·0 Å resolution

Umland, Timothy C.; Swaminathan, Subramanyam; Furey, W.; Singh, Gurmukh; Pletcher, J.; Sax, M.

doi:10.1016/0022-2836(92)91006-b

Cited by 40 publications

(30 citation statements)

References 43 publications

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“…The results of these and other studies [27,28] showed that rabbit and human UGs are indistinguishable proteins both structurally [29] and functionally [23].…”

mentioning

confidence: 76%

“…However, the results of our binding assays fail to support this assumption in that only H-tag-rhUG but not rhUG without the H-tag can bind 45 Ca þþ . Furthermore, the results of structural studies with rabbit and human UGs did not reveal a Ca þþ -binding motif in this protein [25][26][27][28][29]. Thus, the results of our studies demonstrating Lys 43 and Asp 46 in rhUG to be crucial for its sPLA 2 -inhibitory activity and that rhUG without the H-tag fails to bind Ca þþ , it is highly unlikely that sequestration of Ca þþ by this protein is one of the mechanisms by which hUG inhibits sPLA 2 activity.…”

Section: Discussionmentioning

confidence: 99%

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Lys 43 and Asp 46 in α-helix 3 of uteroglobin are essential for its phospholipase A2 inhibitory activity

Chowdhury¹,

Mantile-Selvaggi²,

Miele³

et al. 2002

Biochemical and Biophysical Research Communications

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Uteroglobin (UG) is an anti-inflammatory, secreted protein with soluble phospholipase A 2 (sPLA 2 )-inhibitory activity. However, the mechanism by which UG inhibits sPLA 2 activity is unknown. UG is a homodimer in which each of the 70-amino acid subunits forms four a-helices. We previously reported that sPLA 2 -inhibitory activity of UG may reside in a segment of a-helix 3 that is exposed to the solvent. In addition, it has been suggested that UG may inhibit sPLA 2 activity by binding and sequestering Ca þþ , essential for sPLA 2 activation. By site-specific mutation, we demonstrate here that Lys 43 Glu, Asp 46 Lys or a combination of the two mutations in the full-length, recombinant human UG (rhUG) abrogates its sPLA 2 -inhibitory activity. We demonstrate further that recombinant UG does not bind Ca þþ although when it is expressed with histidine-tag (H-tag) it is capable of binding Ca þþ . Taken together our results show that: (i) Lys 43 and Asp 46 in rhUG are critical residues for the sPLA 2 -inhibitory activity of UG and (ii) Ca þþ -sequestration by rhUG is not likely to be one of the mechanisms responsible for its sPLA 2 -inhibitory activity. Ó 2002 Elsevier Science (USA). All rights reserved.

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“…The results of these and other studies [27,28] showed that rabbit and human UGs are indistinguishable proteins both structurally [29] and functionally [23].…”

mentioning

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Lys 43 and Asp 46 in α-helix 3 of uteroglobin are essential for its phospholipase A2 inhibitory activity

Chowdhury¹,

Mantile-Selvaggi²,

Miele³

et al. 2002

Biochemical and Biophysical Research Communications

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“…One of the major secretory products of human pulmonary Clara cells is the 10-kD protein (cc 1OkD)' ( 1 ), also known as 1 7-kD protein (2). This protein is a homodimer consisting of 70 amino acid subunits connected by two disulfide bonds (3).…”

Section: Introductionmentioning

confidence: 99%

Tissue-specific expression of the gene coding for human Clara cell 10-kD protein, a phospholipase A2-inhibitory protein.

Peri

Cordella‐Miele²,

Miele³

et al. 1993

J. Clin. Invest.

119

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“…The structure of PCB-BP/uteroglobin has been extensively studied and three-dimensional structures based on X-ray diffraction data exist for the rabbit, rat and human protein [13][14][15]. Although several hypotheses exist regarding the physiological role of PCB-BP/uteroglobin (recently reviewed by Miele Abbreviations: BAL, bronchoalveolar lavage; BL, bronchial lavage; CCI0, Clara cell 10-kDa protein; CCSP, Clara cell secretory protein; FPLC, fast protein liquid chromatography; Mr, approximate molecular weight; PAGE, polyacrylamide gel electrophoresis; PCB, polychlorinated biphenyl; PCB-BP, polychlorinated biphenyl-binding protein; PtdChol, phosphatidyl choline; PtdEtn, phosphatidyl ethanolamine; Ptdlns, phosphatidyl inositol; PtdSer, phosphatidyl serine; sPLA2, secretory phospholipase A 2.…”

Section: Introductionmentioning

confidence: 99%

Calcium‐dependent binding of uteroglobin (PCB‐BP/CCSP) to negatively charged phospholipid liposomes

1995

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To investigate interactions between the polyehlorinated biphenyl-binding protein uteroglobin and phospholipids, we used a liposome-pelletting assay. PCB-BPluteroglobin bound to liposomes made from negatively charged phospholipids (PtdSer and Ptdlns) in the presence of 5 mM calcium. No binding to liposomes made from phospholipids without net charge (PtdChol and PtdEtn) was observed, nor could we detect binding in the absence of calcium or when magnesium was substituted for calcium. This suggests that PCB-BPluteroglobin can bind to phospholipids in vivo and may have a role in the phospbolipid homeostasis of the airway and/or secretory pathway of the Clara cell.

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Refined structure of rat Clara cell 17 kDa protein at 3·0 Å resolution

Cited by 40 publications

References 43 publications

Lys 43 and Asp 46 in α-helix 3 of uteroglobin are essential for its phospholipase A2 inhibitory activity

Lys 43 and Asp 46 in α-helix 3 of uteroglobin are essential for its phospholipase A2 inhibitory activity

Tissue-specific expression of the gene coding for human Clara cell 10-kD protein, a phospholipase A2-inhibitory protein.

Calcium‐dependent binding of uteroglobin (PCB‐BP/CCSP) to negatively charged phospholipid liposomes

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