Atomic mutations at the single tryptophan residue of human recombinant annexin V: effects on structure, stability, and activity.


Abstract

The single tryptophan residue (Trp187) of human recombinant annexin V, containing 320 residues and 5328 atoms, was replaced with three different isosteric analogues where hydrogen atoms at positions 4, 5, and 6 in the indole ring were exchanged with fluorine. Such single atom exchanges of H --> F represent atomic mutations that result in slightly increased covalent bond lengths and inverted polarities in the residue side-chain structure. These minimal changes in the local geometry do not affect the secondary and tertiary structures of the mutants, which were identical to those of wild-type protein in the crystal form. But the mutants exhibit significant differences in stability, folding cooperativity, biological activity, and fluorescence properties if compared to the wild-type protein. These rather large global effects, resulting from the minimal local changes, have to be attributed either to the relatively strong changes in polar interactions of the indole ring or to differences in the van der Waals radii or to a combination of both facts. The changes in local geometry that are below resolution of protein X-ray crystallographic studies are probably of secondary importance in comparison to the strong electronegativity introduced by the fluorine atom. Correspondingly, these types of mutations provide an interesting approach to study cooperative functions of integrated residues and modulation of particular physicochemical properties, in the present case of electronegativity, in a uniquely structured and hierarchically organized protein molecule. Study holds ProTherm entries: 6564 Extra Details: tryptophan; fluorine; tertiary structure; van der Waals radii;,local geometry; physicochemical property

Submission Details

ID: LXbmRM4g3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:32 p.m.

Version: 1

Publication Details
Minks C;Huber R;Moroder L;Budisa N,Biochemistry (1999) Atomic mutations at the single tryptophan residue of human recombinant annexin V: effects on structure, stability, and activity. PMID:10451359
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
2IE7 2007-02-06 1.75 Annexin V under 2.0 MPa pressure of nitrous oxide
2IE6 2007-02-06 1.83 Annexin V under 2.0 MPa pressure of xenon
2RAN 1994-11-30 1.89 RAT ANNEXIN V CRYSTAL STRUCTURE: CA2+-INDUCED CONFORMATIONAL CHANGES
1A8B 1998-06-17 1.9 RAT ANNEXIN V COMPLEXED WITH GLYCEROPHOSPHOETHANOLAMINE
1G5N 2001-06-06 1.9 ANNEXIN V COMPLEX WITH HEPARIN OLIGOSACCHARIDES
1ANX 1994-12-20 1.9 THE CRYSTAL STRUCTURE OF A NEW HIGH-CALCIUM FORM OF ANNEXIN V
1A8A 1998-06-17 1.9 RAT ANNEXIN V COMPLEXED WITH GLYCEROPHOSPHOSERINE
1BC3 1998-11-25 1.95 RECOMBINANT RAT ANNEXIN V, TRIPLE MUTANT (T72K, S144K, S228K)
1BCY 1998-11-25 1.95 RECOMBINANT RAT ANNEXIN V, T72K MUTANT
1BC0 1998-11-25 2.0 RECOMBINANT RAT ANNEXIN V, W185A MUTANT
1HVF 1995-03-31 2.0 STRUCTURAL AND ELECTROPHYSIOLOGICAL ANALYSIS OF ANNEXIN V MUTANTS. MUTAGENESIS OF HUMAN ANNEXIN V, AN IN VITRO VOLTAGE-GATED CALCIUM CHANNEL, PROVIDES INFORMATION ABOUT THE STRUCTURAL FEATURES OF THE ION PATHWAY, THE VOLTAGE SENSOR AND THE ION SELECTIVITY FILTER
1HVD 1995-03-31 2.0 STRUCTURAL AND ELECTROPHYSIOLOGICAL ANALYSIS OF ANNEXIN V MUTANTS. MUTAGENESIS OF HUMAN ANNEXIN V, AN IN VITRO VOLTAGE-GATED CALCIUM CHANNEL, PROVIDES INFORMATION ABOUT THE STRUCTURAL FEATURES OF THE ION PATHWAY, THE VOLTAGE SENSOR AND THE ION SELECTIVITY FILTER
1BC1 1998-11-25 2.05 RECOMBINANT RAT ANNEXIN V, QUADRUPLE MUTANT (T72K, S144K, S228K, S303K)
1N42 2003-02-04 2.1 Crystal Structure of Annexin V R149E Mutant
1N41 2003-02-04 2.1 Crystal Structure of Annexin V K27E Mutant
1BCW 1998-11-25 2.1 RECOMBINANT RAT ANNEXIN V, T72A MUTANT
1BCZ 1998-11-25 2.2 RECOMBINANT RAT ANNEXIN V, T72S MUTANT
2H0M 2007-06-19 2.26 Structure of a Mutant of Rat Annexin A5
1HVE 1995-03-31 2.3 STRUCTURAL AND ELECTROPHYSIOLOGICAL ANALYSIS OF ANNEXIN V MUTANTS. MUTAGENESIS OF HUMAN ANNEXIN V, AN IN VITRO VOLTAGE-GATED CALCIUM CHANNEL, PROVIDES INFORMATION ABOUT THE STRUCTURAL FEATURES OF THE ION PATHWAY, THE VOLTAGE SENSOR AND THE ION SELECTIVITY FILTER
2XO3 2011-08-03 2.3 Human Annexin V with incorporated Methionine analogue Homopropargylglycine
1AVH 1994-01-31 2.3 CRYSTAL AND MOLECULAR STRUCTURE OF HUMAN ANNEXIN V AFTER REFINEMENT. IMPLICATIONS FOR STRUCTURE, MEMBRANE BINDING AND ION CHANNEL FORMATION OF THE ANNEXIN FAMILY OF PROTEINS
1AVR 1994-01-31 2.3 CRYSTAL AND MOLECULAR STRUCTURE OF HUMAN ANNEXIN V AFTER REFINEMENT. IMPLICATIONS FOR STRUCTURE, MEMBRANE BINDING AND ION CHANNEL FORMATION OF THE ANNEXIN FAMILY OF PROTEINS
1ANW 1994-12-20 2.4 THE EFFECT OF METAL BINDING ON THE STRUCTURE OF ANNEXIN V AND IMPLICATIONS FOR MEMBRANE BINDING
1SAV 1998-05-27 2.5 HUMAN ANNEXIN V WITH PROLINE SUBSTITUTION BY THIOPROLINE
2H0L 2007-06-19 2.59 Crystal Structure of a Mutant of Rat Annexin A5
2H0K 2007-06-19 2.76 Crystal Structure of a Mutant of Rat Annexin A5
2XO2 2011-08-03 2.8 Human Annexin V with incorporated Methionine analogue Azidohomoalanine
1N44 2003-02-04 3.0 Crystal Structure of Annexin V R23E Mutant
1HAK 1999-02-16 3.0 CRYSTAL STRUCTURE OF RECOMBINANT HUMAN PLACENTAL ANNEXIN V COMPLEXED WITH K-201 AS A CALCIUM CHANNEL ACTIVITY INHIBITOR
1HVG 1995-03-31 3.0 STRUCTURAL AND ELECTROPHYSIOLOGICAL ANALYSIS OF ANNEXIN V MUTANTS. MUTAGENESIS OF HUMAN ANNEXIN V, AN IN VITRO VOLTAGE-GATED CALCIUM CHANNEL, PROVIDES INFORMATION ABOUT THE STRUCTURAL FEATURES OF THE ION PATHWAY, THE VOLTAGE SENSOR AND THE ION SELECTIVITY FILTER

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
91.8 Annexin A5 P14668 ANXA5_RAT
93.7 Annexin A5 P48036 ANXA5_MOUSE
95.9 Annexin A5 P81287 ANXA5_BOVIN
98.4 Annexin A5 Q4R4H7 ANXA5_MACFA
100.0 Annexin A5 Q5R1W0 ANXA5_PANTR
100.0 Annexin A5 P08758 ANXA5_HUMAN